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Patrick C. Y. Woo - One of the best experts on this subject based on the ideXlab platform.
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novel partitivirus enhances virulence of and causes aberrant gene expression in talaromyces marneffei
Mbio, 2018Co-Authors: Susanna K. P. Lau, Franklin W. N. Chow, Rachel Y. Y. Fan, James J. Cai, Kwok-yung Yuen, Patrick C. Y. WooAbstract:Talaromyces marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in Southeast Asia. We report the discovery of a novel partitivirus, Talaromyces marneffeipartitivirus-1 (TmPV1). TmPV1 was detected in 7 (12.7%) of 55 clinical T. marneffei isolates. Complete genome sequencing of the seven TmPV1 isolates revealed two double-stranded RNA (dsRNA) segments encoding RNA-dependent RNA polymerase (RdRp) and capsid protein, respectively. Phylogenetic analysis showed that TmPV1 occupied a distinct clade among the members of the genus Gammapartitivirus Transmission electron microscopy confirmed the presence of isometric, nonenveloped viral particles of 30 to 45 nm in diameter, compatible with partitiviruses, in TmPV1-infected T. marneffei Quantitative reverse transcription-PCR (qRT-PCR) demonstrated higher viral load of TmPV1 in the yeast phase than in the mycelial phase of T. marneffei Two virus-free isolates, PM1 and PM41, were successfully infected by purified TmPV1 using protoplast transfection. Mice challenged with TmPV1-infected T. marneffei isolates showed significantly shortened survival time (P < 0.0001) and higher fungal burden in organs than mice challenged with isogenic TmPV1-free isolates. Transcriptomic analysis showed that TmPV1 causes aberrant expression of various genes in T. marneffei, with upregulation of potential virulence factors and suppression of RNA interference (RNAi)-related genes. This is the first report of a mycovirus in a thermally Dimorphic Fungus. Further studies are required to ascertain the mechanism whereby TmPV1 enhances the virulence of T. marneffei in mice and the potential role of RNAi-related genes in antiviral defense in T. marneffeiIMPORTANCETalaromyces marneffei (formerly Penicillium marneffei) is the most important thermal Dimorphic Fungus in Southeast Asia, causing highly fatal systemic penicilliosis in HIV-infected and immunocompromised patients. We discovered a novel mycovirus, TmPV1, in seven clinical isolates of T. marneffei TmPV1 belongs to the genus Gammapartitivirus of the family Partitiviridae We showed that TmPV1 enhanced the virulence of T. marneffei in mice, with shortened survival time and higher fungal burden in the organs of mice challenged with TmPV1-infected T. marneffei isolates than in those of mice challenged with virus-free isogenic isolates. Transcriptomics analysis showed that TmPV1 altered the expression of genes involved in various cellular processes in T. marneffei, with upregulation of potential virulence factors and suppression of RNAi machinery which may be involved in antiviral defense. This is the first report of a mycovirus in a thermal Dimorphic Fungus. The present results offer insights into mycovirus-Fungus interactions and pathogenesis of thermal Dimorphic fungi.
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Novel Partitivirus Enhances Virulence of and Causes Aberrant Gene Expression in Talaromyces marneffei
American Society for Microbiology, 2018Co-Authors: Susanna K. P. Lau, Franklin W. N. Chow, Rachel Y. Y. Fan, James J. Cai, Kwok-yung Yuen, Patrick C. Y. WooAbstract:Talaromyces marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in Southeast Asia. We report the discovery of a novel partitivirus, Talaromyces marneffeipartitivirus-1 (TmPV1). TmPV1 was detected in 7 (12.7%) of 55 clinical T. marneffei isolates. Complete genome sequencing of the seven TmPV1 isolates revealed two double-stranded RNA (dsRNA) segments encoding RNA-dependent RNA polymerase (RdRp) and capsid protein, respectively. Phylogenetic analysis showed that TmPV1 occupied a distinct clade among the members of the genus Gammapartitivirus. Transmission electron microscopy confirmed the presence of isometric, nonenveloped viral particles of 30 to 45 nm in diameter, compatible with partitiviruses, in TmPV1-infected T. marneffei. Quantitative reverse transcription-PCR (qRT-PCR) demonstrated higher viral load of TmPV1 in the yeast phase than in the mycelial phase of T. marneffei. Two virus-free isolates, PM1 and PM41, were successfully infected by purified TmPV1 using protoplast transfection. Mice challenged with TmPV1-infected T. marneffei isolates showed significantly shortened survival time (P < 0.0001) and higher fungal burden in organs than mice challenged with isogenic TmPV1-free isolates. Transcriptomic analysis showed that TmPV1 causes aberrant expression of various genes in T. marneffei, with upregulation of potential virulence factors and suppression of RNA interference (RNAi)-related genes. This is the first report of a mycovirus in a thermally Dimorphic Fungus. Further studies are required to ascertain the mechanism whereby TmPV1 enhances the virulence of T. marneffei in mice and the potential role of RNAi-related genes in antiviral defense in T. marneffei.Talaromyces marneffei (formerly Penicillium marneffei) is the most important thermal Dimorphic Fungus in Southeast Asia, causing highly fatal systemic penicilliosis in HIV-infected and immunocompromised patients. We discovered a novel mycovirus, TmPV1, in seven clinical isolates of T. marneffei. TmPV1 belongs to the genus Gammapartitivirus of the family Partitiviridae. We showed that TmPV1 enhanced the virulence of T. marneffei in mice, with shortened survival time and higher fungal burden in the organs of mice challenged with TmPV1-infected T. marneffei isolates than in those of mice challenged with virus-free isogenic isolates. Transcriptomics analysis showed that TmPV1 altered the expression of genes involved in various cellular processes in T. marneffei, with upregulation of potential virulence factors and suppression of RNAi machinery which may be involved in antiviral defense. This is the first report of a mycovirus in a thermal Dimorphic Fungus. The present results offer insights into mycovirus-Fungus interactions and pathogenesis of thermal Dimorphic fungi
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Functional Analysis of atfA Gene to Stress Response in Pathogenic Thermal Dimorphic Fungus Penicillium marneffei
2016Co-Authors: Panjaphorn Nimmanee, Patrick C. Y. Woo, Pramote Vanittanakom, Sirida Youngchim, Nongnuch VanittanakomAbstract:Penicillium marneffei, the pathogenic thermal Dimorphic Fungus is a causative agent of a fatal systemic disease, penicilliosis marneffei, in immunocompromised patients especially HIV patients. For growth and survival, this Fungus has to adapt to environmental stresses outside and inside host cells and this adaptation requires stress signaling pathways and regulation of gene expression under various kinds of stresses. In this report, P. marneffei activating transcription factor (atfA) gene encoding bZip-type transcription factor was characterized. To determine functions of this gene, atfA isogenic mutant strain was constructed using the modified split marker recombination method. The phenotypes and susceptibility to varieties of stresses including osmotic, oxidative, heat, UV, cell wall and cell membrane stresses of the mutant strain were compared with the wild type and the atfA complemented strains. Results demonstrated that the mRNA expression level of P. marneffei atfA gene increased under heat stress at 42uC. The atfA mutant was more sensitive to sodium dodecyl sulphate, amphotericin B and tert-butyl hydroperoxide than the wild type and complemented strains but not hydrogen peroxide, menadione, NaCl, sorbitol, calcofluor white, itraconazole, UV stresses and heat stress at 39uC. In addition, recovery of atfA mutant conidia after mouse and human macrophage infections was significantly decreased compared to those of wild type and complemented strains. These results indicated that the atfA gene was required by P. marneffei under specific stres
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matrix assisted laser desorption ionization time of flight mass spectrometry for rapid identification of mold and yeast cultures of penicillium marneffei
BMC Microbiology, 2016Co-Authors: Susanna K. P. Lau, Clare S K Lam, Antonio H Y Ngan, Wangngai Chow, Dominic N C Tsang, Cindy W S Tse, Taklun Que, Bone S F Tang, Patrick C. Y. WooAbstract:Background Penicillium marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in HIV-infected and other immunocompromised patients in Southeast Asia. However, laboratory diagnosis of penicilliosis, which relies on microscopic morphology and mycelial-to-yeast conversion, is time-consuming and expertise-dependent, thus delaying diagnosis and treatment. Although matrix -assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is useful for identification of various medically important fungi, its performance for identification of P. marneffei is less clear.
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Mp1p Is a Virulence Factor in Talaromyces (Penicillium) marneffei.
Public Library of Science (PLoS), 2016Co-Authors: Patrick C. Y. Woo, Susanna K. P. Lau, Ken T. K. Chong, Candy C Y Lau, Edward T K Tung, Fengjuan Yang, Hongmin Zhang, Jian-pao Cai, Rex K H Au-yeungAbstract:Talaromyces marneffei is an opportunistic Dimorphic Fungus prevalent in Southeast Asia. We previously demonstrated that Mp1p is an immunogenic surface and secretory mannoprotein of T. marneffei. Since Mp1p is a surface protein that can generate protective immunity, we hypothesized that Mp1p and/or its homologs are virulence factors.We examined the pathogenic roles of Mp1p and its homologs in a mouse model. All mice died 21 and 30 days after challenge with wild-type T. marneffei PM1 and MP1 complemented mutant respectively. None of the mice died 60 days after challenge with MP1 knockout mutant (P
Susanna K. P. Lau - One of the best experts on this subject based on the ideXlab platform.
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novel partitivirus enhances virulence of and causes aberrant gene expression in talaromyces marneffei
Mbio, 2018Co-Authors: Susanna K. P. Lau, Franklin W. N. Chow, Rachel Y. Y. Fan, James J. Cai, Kwok-yung Yuen, Patrick C. Y. WooAbstract:Talaromyces marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in Southeast Asia. We report the discovery of a novel partitivirus, Talaromyces marneffeipartitivirus-1 (TmPV1). TmPV1 was detected in 7 (12.7%) of 55 clinical T. marneffei isolates. Complete genome sequencing of the seven TmPV1 isolates revealed two double-stranded RNA (dsRNA) segments encoding RNA-dependent RNA polymerase (RdRp) and capsid protein, respectively. Phylogenetic analysis showed that TmPV1 occupied a distinct clade among the members of the genus Gammapartitivirus Transmission electron microscopy confirmed the presence of isometric, nonenveloped viral particles of 30 to 45 nm in diameter, compatible with partitiviruses, in TmPV1-infected T. marneffei Quantitative reverse transcription-PCR (qRT-PCR) demonstrated higher viral load of TmPV1 in the yeast phase than in the mycelial phase of T. marneffei Two virus-free isolates, PM1 and PM41, were successfully infected by purified TmPV1 using protoplast transfection. Mice challenged with TmPV1-infected T. marneffei isolates showed significantly shortened survival time (P < 0.0001) and higher fungal burden in organs than mice challenged with isogenic TmPV1-free isolates. Transcriptomic analysis showed that TmPV1 causes aberrant expression of various genes in T. marneffei, with upregulation of potential virulence factors and suppression of RNA interference (RNAi)-related genes. This is the first report of a mycovirus in a thermally Dimorphic Fungus. Further studies are required to ascertain the mechanism whereby TmPV1 enhances the virulence of T. marneffei in mice and the potential role of RNAi-related genes in antiviral defense in T. marneffeiIMPORTANCETalaromyces marneffei (formerly Penicillium marneffei) is the most important thermal Dimorphic Fungus in Southeast Asia, causing highly fatal systemic penicilliosis in HIV-infected and immunocompromised patients. We discovered a novel mycovirus, TmPV1, in seven clinical isolates of T. marneffei TmPV1 belongs to the genus Gammapartitivirus of the family Partitiviridae We showed that TmPV1 enhanced the virulence of T. marneffei in mice, with shortened survival time and higher fungal burden in the organs of mice challenged with TmPV1-infected T. marneffei isolates than in those of mice challenged with virus-free isogenic isolates. Transcriptomics analysis showed that TmPV1 altered the expression of genes involved in various cellular processes in T. marneffei, with upregulation of potential virulence factors and suppression of RNAi machinery which may be involved in antiviral defense. This is the first report of a mycovirus in a thermal Dimorphic Fungus. The present results offer insights into mycovirus-Fungus interactions and pathogenesis of thermal Dimorphic fungi.
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Novel Partitivirus Enhances Virulence of and Causes Aberrant Gene Expression in Talaromyces marneffei
American Society for Microbiology, 2018Co-Authors: Susanna K. P. Lau, Franklin W. N. Chow, Rachel Y. Y. Fan, James J. Cai, Kwok-yung Yuen, Patrick C. Y. WooAbstract:Talaromyces marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in Southeast Asia. We report the discovery of a novel partitivirus, Talaromyces marneffeipartitivirus-1 (TmPV1). TmPV1 was detected in 7 (12.7%) of 55 clinical T. marneffei isolates. Complete genome sequencing of the seven TmPV1 isolates revealed two double-stranded RNA (dsRNA) segments encoding RNA-dependent RNA polymerase (RdRp) and capsid protein, respectively. Phylogenetic analysis showed that TmPV1 occupied a distinct clade among the members of the genus Gammapartitivirus. Transmission electron microscopy confirmed the presence of isometric, nonenveloped viral particles of 30 to 45 nm in diameter, compatible with partitiviruses, in TmPV1-infected T. marneffei. Quantitative reverse transcription-PCR (qRT-PCR) demonstrated higher viral load of TmPV1 in the yeast phase than in the mycelial phase of T. marneffei. Two virus-free isolates, PM1 and PM41, were successfully infected by purified TmPV1 using protoplast transfection. Mice challenged with TmPV1-infected T. marneffei isolates showed significantly shortened survival time (P < 0.0001) and higher fungal burden in organs than mice challenged with isogenic TmPV1-free isolates. Transcriptomic analysis showed that TmPV1 causes aberrant expression of various genes in T. marneffei, with upregulation of potential virulence factors and suppression of RNA interference (RNAi)-related genes. This is the first report of a mycovirus in a thermally Dimorphic Fungus. Further studies are required to ascertain the mechanism whereby TmPV1 enhances the virulence of T. marneffei in mice and the potential role of RNAi-related genes in antiviral defense in T. marneffei.Talaromyces marneffei (formerly Penicillium marneffei) is the most important thermal Dimorphic Fungus in Southeast Asia, causing highly fatal systemic penicilliosis in HIV-infected and immunocompromised patients. We discovered a novel mycovirus, TmPV1, in seven clinical isolates of T. marneffei. TmPV1 belongs to the genus Gammapartitivirus of the family Partitiviridae. We showed that TmPV1 enhanced the virulence of T. marneffei in mice, with shortened survival time and higher fungal burden in the organs of mice challenged with TmPV1-infected T. marneffei isolates than in those of mice challenged with virus-free isogenic isolates. Transcriptomics analysis showed that TmPV1 altered the expression of genes involved in various cellular processes in T. marneffei, with upregulation of potential virulence factors and suppression of RNAi machinery which may be involved in antiviral defense. This is the first report of a mycovirus in a thermal Dimorphic Fungus. The present results offer insights into mycovirus-Fungus interactions and pathogenesis of thermal Dimorphic fungi
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matrix assisted laser desorption ionization time of flight mass spectrometry for rapid identification of mold and yeast cultures of penicillium marneffei
BMC Microbiology, 2016Co-Authors: Susanna K. P. Lau, Clare S K Lam, Antonio H Y Ngan, Wangngai Chow, Dominic N C Tsang, Cindy W S Tse, Taklun Que, Bone S F Tang, Patrick C. Y. WooAbstract:Background Penicillium marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in HIV-infected and other immunocompromised patients in Southeast Asia. However, laboratory diagnosis of penicilliosis, which relies on microscopic morphology and mycelial-to-yeast conversion, is time-consuming and expertise-dependent, thus delaying diagnosis and treatment. Although matrix -assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is useful for identification of various medically important fungi, its performance for identification of P. marneffei is less clear.
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Mp1p Is a Virulence Factor in Talaromyces (Penicillium) marneffei.
Public Library of Science (PLoS), 2016Co-Authors: Patrick C. Y. Woo, Susanna K. P. Lau, Ken T. K. Chong, Candy C Y Lau, Edward T K Tung, Fengjuan Yang, Hongmin Zhang, Jian-pao Cai, Rex K H Au-yeungAbstract:Talaromyces marneffei is an opportunistic Dimorphic Fungus prevalent in Southeast Asia. We previously demonstrated that Mp1p is an immunogenic surface and secretory mannoprotein of T. marneffei. Since Mp1p is a surface protein that can generate protective immunity, we hypothesized that Mp1p and/or its homologs are virulence factors.We examined the pathogenic roles of Mp1p and its homologs in a mouse model. All mice died 21 and 30 days after challenge with wild-type T. marneffei PM1 and MP1 complemented mutant respectively. None of the mice died 60 days after challenge with MP1 knockout mutant (P
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Mp1p Is a Virulence Factor in Talaromyces (Penicillium) marneffei
2016Co-Authors: Patrick C. Y. Woo, Susanna K. P. Lau, Ken T. K. Chong, Candy C Y Lau, Edward T K Tung, Fengjuan Yang, Hongmin Zhang, Jian-pao Cai, Rex K H Au-yeungAbstract:BackgroundTalaromyces marneffei is an opportunistic Dimorphic Fungus prevalent in Southeast Asia. We previously demonstrated that Mp1p is an immunogenic surface and secretory mannoprotein of T. marneffei. Since Mp1p is a surface protein that can generate protective immunity, we hypothesized that Mp1p and/or its homologs are virulence factors.Methodology/Principal FindingsWe examined the pathogenic roles of Mp1p and its homologs in a mouse model. All mice died 21 and 30 days after challenge with wild-type T. marneffei PM1 and MP1 complemented mutant respectively. None of the mice died 60 days after challenge with MP1 knockout mutant (P
Kwok-yung Yuen - One of the best experts on this subject based on the ideXlab platform.
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novel partitivirus enhances virulence of and causes aberrant gene expression in talaromyces marneffei
Mbio, 2018Co-Authors: Susanna K. P. Lau, Franklin W. N. Chow, Rachel Y. Y. Fan, James J. Cai, Kwok-yung Yuen, Patrick C. Y. WooAbstract:Talaromyces marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in Southeast Asia. We report the discovery of a novel partitivirus, Talaromyces marneffeipartitivirus-1 (TmPV1). TmPV1 was detected in 7 (12.7%) of 55 clinical T. marneffei isolates. Complete genome sequencing of the seven TmPV1 isolates revealed two double-stranded RNA (dsRNA) segments encoding RNA-dependent RNA polymerase (RdRp) and capsid protein, respectively. Phylogenetic analysis showed that TmPV1 occupied a distinct clade among the members of the genus Gammapartitivirus Transmission electron microscopy confirmed the presence of isometric, nonenveloped viral particles of 30 to 45 nm in diameter, compatible with partitiviruses, in TmPV1-infected T. marneffei Quantitative reverse transcription-PCR (qRT-PCR) demonstrated higher viral load of TmPV1 in the yeast phase than in the mycelial phase of T. marneffei Two virus-free isolates, PM1 and PM41, were successfully infected by purified TmPV1 using protoplast transfection. Mice challenged with TmPV1-infected T. marneffei isolates showed significantly shortened survival time (P < 0.0001) and higher fungal burden in organs than mice challenged with isogenic TmPV1-free isolates. Transcriptomic analysis showed that TmPV1 causes aberrant expression of various genes in T. marneffei, with upregulation of potential virulence factors and suppression of RNA interference (RNAi)-related genes. This is the first report of a mycovirus in a thermally Dimorphic Fungus. Further studies are required to ascertain the mechanism whereby TmPV1 enhances the virulence of T. marneffei in mice and the potential role of RNAi-related genes in antiviral defense in T. marneffeiIMPORTANCETalaromyces marneffei (formerly Penicillium marneffei) is the most important thermal Dimorphic Fungus in Southeast Asia, causing highly fatal systemic penicilliosis in HIV-infected and immunocompromised patients. We discovered a novel mycovirus, TmPV1, in seven clinical isolates of T. marneffei TmPV1 belongs to the genus Gammapartitivirus of the family Partitiviridae We showed that TmPV1 enhanced the virulence of T. marneffei in mice, with shortened survival time and higher fungal burden in the organs of mice challenged with TmPV1-infected T. marneffei isolates than in those of mice challenged with virus-free isogenic isolates. Transcriptomics analysis showed that TmPV1 altered the expression of genes involved in various cellular processes in T. marneffei, with upregulation of potential virulence factors and suppression of RNAi machinery which may be involved in antiviral defense. This is the first report of a mycovirus in a thermal Dimorphic Fungus. The present results offer insights into mycovirus-Fungus interactions and pathogenesis of thermal Dimorphic fungi.
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Novel Partitivirus Enhances Virulence of and Causes Aberrant Gene Expression in Talaromyces marneffei
American Society for Microbiology, 2018Co-Authors: Susanna K. P. Lau, Franklin W. N. Chow, Rachel Y. Y. Fan, James J. Cai, Kwok-yung Yuen, Patrick C. Y. WooAbstract:Talaromyces marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in Southeast Asia. We report the discovery of a novel partitivirus, Talaromyces marneffeipartitivirus-1 (TmPV1). TmPV1 was detected in 7 (12.7%) of 55 clinical T. marneffei isolates. Complete genome sequencing of the seven TmPV1 isolates revealed two double-stranded RNA (dsRNA) segments encoding RNA-dependent RNA polymerase (RdRp) and capsid protein, respectively. Phylogenetic analysis showed that TmPV1 occupied a distinct clade among the members of the genus Gammapartitivirus. Transmission electron microscopy confirmed the presence of isometric, nonenveloped viral particles of 30 to 45 nm in diameter, compatible with partitiviruses, in TmPV1-infected T. marneffei. Quantitative reverse transcription-PCR (qRT-PCR) demonstrated higher viral load of TmPV1 in the yeast phase than in the mycelial phase of T. marneffei. Two virus-free isolates, PM1 and PM41, were successfully infected by purified TmPV1 using protoplast transfection. Mice challenged with TmPV1-infected T. marneffei isolates showed significantly shortened survival time (P < 0.0001) and higher fungal burden in organs than mice challenged with isogenic TmPV1-free isolates. Transcriptomic analysis showed that TmPV1 causes aberrant expression of various genes in T. marneffei, with upregulation of potential virulence factors and suppression of RNA interference (RNAi)-related genes. This is the first report of a mycovirus in a thermally Dimorphic Fungus. Further studies are required to ascertain the mechanism whereby TmPV1 enhances the virulence of T. marneffei in mice and the potential role of RNAi-related genes in antiviral defense in T. marneffei.Talaromyces marneffei (formerly Penicillium marneffei) is the most important thermal Dimorphic Fungus in Southeast Asia, causing highly fatal systemic penicilliosis in HIV-infected and immunocompromised patients. We discovered a novel mycovirus, TmPV1, in seven clinical isolates of T. marneffei. TmPV1 belongs to the genus Gammapartitivirus of the family Partitiviridae. We showed that TmPV1 enhanced the virulence of T. marneffei in mice, with shortened survival time and higher fungal burden in the organs of mice challenged with TmPV1-infected T. marneffei isolates than in those of mice challenged with virus-free isogenic isolates. Transcriptomics analysis showed that TmPV1 altered the expression of genes involved in various cellular processes in T. marneffei, with upregulation of potential virulence factors and suppression of RNAi machinery which may be involved in antiviral defense. This is the first report of a mycovirus in a thermal Dimorphic Fungus. The present results offer insights into mycovirus-Fungus interactions and pathogenesis of thermal Dimorphic fungi
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proteome profiling of the Dimorphic Fungus penicillium marneffei extracellular proteins and identification of glyceraldehyde 3 phosphate dehydrogenase as an important adhesion factor for conidial attachment
FEBS Journal, 2013Co-Authors: Susanna K. P. Lau, Kwok-yung Yuen, Herman Tse, Joanna S Y Chan, Anna C Zhou, Shirly O T Curreem, Candy C Y Lau, Patrick C. Y. WooAbstract:Despite being the most important thermal Dimorphic Fungus causing systemic mycosis in Southeast Asia, the pathogenic mechanisms of Penicillium marneffei remain largely unknown. By comparing the extracellular proteomes of P. marneffei in mycelial and yeast phases, we identified 12 differentially expressed proteins among which glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and heat shock protein 60 (HSP60) were found to be upregulated in mycelial and yeast phases respectively. Based on previous findings in other pathogens, we hypothesized that these two extracellular proteins may be involved in adherence during P. marneffei–host interaction. Using inhibition assays with recombinant GAPDH (rGAPDH) proteins and anti-rGAPDH sera, we demonstrated that adhesion of P. marneffei conidia to fibronectin and laminin was inhibited by rGAPDH or rabbit anti-rGAPDH serum in a dose-dependent manner. Similarly, a dose-dependent inhibition of conidial adherence to A549 pneumocytes by rGAPDH or rabbit anti-rGAPDH serum was observed, suggesting that P. marneffei GAPDH can mediate binding of conidia to human extracellular matrix proteins and pneumocytes. However, HSP60 did not exhibit similar inhibition on conidia adherence, and neither GAPDH norHSP60 exhibited inhibition on adherence to J774 or THP-1 macrophage cell lines. This report demonstrates GAPDH as an adherence factor in P. marneffei by mediating conidia adherence to host bronchoalveolar epithelium during the early establishment phase of infection.
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identification of microrna like rnas in mycelial and yeast phases of the thermal Dimorphic Fungus penicillium marneffei
PLOS Neglected Tropical Diseases, 2013Co-Authors: Susanna K. P. Lau, Patrick C. Y. Woo, Wangngai Chow, Annette Y P Wong, Julian M Y Yeung, Jessie Y J Bao, Na Zhang, Si Lok, Kwok-yung YuenAbstract:Background Penicillium marneffei is the most important thermal Dimorphic Fungus causing systemic mycosis in China and Southeast Asia. While miRNAs are increasingly recognized for their roles in post-transcriptional regulation of gene expression in animals and plants, miRNAs in fungi were less well studied and their potential roles in fungal dimorphism were largely unknown. Based on P. marneffei genome sequence, we hypothesize that miRNA-like RNAs (milRNAs) may be expressed in the Dimorphic Fungus. Methodology/Principal Findings We attempted to identify milRNAs in P. marneffei in both mycelial and yeast phase using high-throughput sequencing technology. Small RNAs were more abundantly expressed in mycelial than yeast phase. Sequence analysis revealed 24 potential milRNA candidates, including 17 candidates in mycelial and seven in yeast phase. Two genes, dcl-1 and dcl-2, encoding putative Dicer-like proteins and the gene, qde-2, encoding Argonaute-like protein, were identified in P. marneffei. Phylogenetic analysis showed that dcl-2 of P. marneffei was more closely related to the homologues in other thermal Dimorphic pathogenic fungi than to Penicillium chrysogenum and Aspergillus spp., suggesting the co-evolution of dcl-2 among the thermal Dimorphic fungi. Moreover, dcl-2 demonstrated higher mRNA expression levels in mycelial than yeast phase by 7 folds (P<0.001). Northern blot analysis confirmed the expression of two milRNAs, PM-milR-M1 and PM-milR-M2, only in mycelial phase. Using dcl-1KO, dcl-2KO, dclDKO and qde-2KO deletion mutants, we showed that the biogenesis of both milRNAs were dependent on dcl-2 but not dcl-1 or qde-2. The mRNA expression levels of three predicted targets of PM-milR-M1 were upregulated in knockdown strain PM-milR-M1KD, supporting regulatory function of milRNAs. Conclusions/Significance Our findings provided the first evidence for differential expression of milRNAs in different growth phases of thermal Dimorphic fungi and shed light on the evolution of fungal proteins involved in milRNA biogenesis and possible role of post-transcriptional control in governing thermal dimorphism.
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penicillium marneffei fungaemia in an allogeneic bone marrow transplant recipient
Bone Marrow Transplantation, 2005Co-Authors: Patrick C. Y. Woo, Susanna K. P. Lau, Samson S. Y. Wong, Ken T. K. Chong, Candy Choiyi Lau, Waiting Hui, Kwok-yung YuenAbstract:Penicillium marneffei is the most important thermal Dimorphic Fungus causing respiratory, skin and systemic mycosis in Southeast Asia.1, 2, 3 About 8% of AIDS patients in Hong Kong are infected with P. marneffei. Clinically, penicilliosis manifests as a systemic febrile illness, which results from intracellular infection of the reticuloendothelial cells by the yeast phase of the Fungus. Besides HIV-positive patients, P. marneffei infections have been reported in other immunocompromised patients, such as renal transplant recipients, patients with systemic lupus erythematosus and patients who are receiving corticosteroid therapy.4, 5 Here, we describe the first case of P. marneffei infection in a bone marrow transplant recipient.
Nongnuch Vanittanakom - One of the best experts on this subject based on the ideXlab platform.
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Functional Analysis of atfA Gene to Stress Response in Pathogenic Thermal Dimorphic Fungus Penicillium marneffei
2016Co-Authors: Panjaphorn Nimmanee, Patrick C. Y. Woo, Pramote Vanittanakom, Sirida Youngchim, Nongnuch VanittanakomAbstract:Penicillium marneffei, the pathogenic thermal Dimorphic Fungus is a causative agent of a fatal systemic disease, penicilliosis marneffei, in immunocompromised patients especially HIV patients. For growth and survival, this Fungus has to adapt to environmental stresses outside and inside host cells and this adaptation requires stress signaling pathways and regulation of gene expression under various kinds of stresses. In this report, P. marneffei activating transcription factor (atfA) gene encoding bZip-type transcription factor was characterized. To determine functions of this gene, atfA isogenic mutant strain was constructed using the modified split marker recombination method. The phenotypes and susceptibility to varieties of stresses including osmotic, oxidative, heat, UV, cell wall and cell membrane stresses of the mutant strain were compared with the wild type and the atfA complemented strains. Results demonstrated that the mRNA expression level of P. marneffei atfA gene increased under heat stress at 42uC. The atfA mutant was more sensitive to sodium dodecyl sulphate, amphotericin B and tert-butyl hydroperoxide than the wild type and complemented strains but not hydrogen peroxide, menadione, NaCl, sorbitol, calcofluor white, itraconazole, UV stresses and heat stress at 39uC. In addition, recovery of atfA mutant conidia after mouse and human macrophage infections was significantly decreased compared to those of wild type and complemented strains. These results indicated that the atfA gene was required by P. marneffei under specific stres
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characterization of saka gene from pathogenic Dimorphic Fungus penicillium marneffei
International Journal of Medical Microbiology, 2015Co-Authors: Panjaphorn Nimmanee, Patrick C. Y. Woo, Aksarakorn Kummasook, Nongnuch VanittanakomAbstract:Eukaryotes utilize stress activated protein kinase (SAPK) pathways to adapt to environmental stress, including heat, osmotic, oxidative or nutrient stresses. Penicillium marneffei (Talaromyces marneffei), the Dimorphic pathogenic Fungus that can cause disseminated mycosis in HIV-infected patients, has to encounter various types of stresses both outside and inside host cells. However, the strategies used by this Fungus in response to these stresses are still unclear. In this report, the stress-activated kinase (sakA) gene of P. marneffei was characterized and the roles of this gene on various stress conditions were studied. The sakA gene deletion mutant was constructed using the split marker method. The phenotypes and sensitivities to varieties of stresses, including osmotic, oxidative, heat and cell wall stresses of the deletion mutant were compared with the wild type and the sakA complemented strains. Results demonstrated that the P. marneffei sakA gene encoded a putative protein containing TXY phosphorylation lip found in the stress high osmolarity glycerol 1 (Hog1)/Spc1/p38 MAPK family, and that this gene was involved not only in tolerance against oxidative and heat stresses, but also played a role in asexual development, chitin deposition, yeast cell generation in vitro and survival inside mouse and human macrophages.
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functional analysis of atfa gene to stress response in pathogenic thermal Dimorphic Fungus penicillium marneffei
PLOS ONE, 2014Co-Authors: Panjaphorn Nimmanee, Patrick C. Y. Woo, Pramote Vanittanakom, Sirida Youngchim, Nongnuch VanittanakomAbstract:Penicillium marneffei, the pathogenic thermal Dimorphic Fungus is a causative agent of a fatal systemic disease, penicilliosis marneffei, in immunocompromised patients especially HIV patients. For growth and survival, this Fungus has to adapt to environmental stresses outside and inside host cells and this adaptation requires stress signaling pathways and regulation of gene expression under various kinds of stresses. In this report, P. marneffei activating transcription factor (atfA) gene encoding bZip-type transcription factor was characterized. To determine functions of this gene, atfA isogenic mutant strain was constructed using the modified split marker recombination method. The phenotypes and susceptibility to varieties of stresses including osmotic, oxidative, heat, UV, cell wall and cell membrane stresses of the mutant strain were compared with the wild type and the atfA complemented strains. Results demonstrated that the mRNA expression level of P. marneffei atfA gene increased under heat stress at 42°C. The atfA mutant was more sensitive to sodium dodecyl sulphate, amphotericin B and tert-butyl hydroperoxide than the wild type and complemented strains but not hydrogen peroxide, menadione, NaCl, sorbitol, calcofluor white, itraconazole, UV stresses and heat stress at 39°C. In addition, recovery of atfA mutant conidia after mouse and human macrophage infections was significantly decreased compared to those of wild type and complemented strains. These results indicated that the atfA gene was required by P. marneffei under specific stress conditions and might be necessary for fighting against host immune cells during the initiation of infection.
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Isolation and Differential Expression of ura5 Gene during Phase Transition and Stress Conditions in a Human Pathogenic Dimorphic Fungus, Penicillium marneffei °“√·¬ ° ·≈– · ¥ßÕÕ°¢Õ߬’π ura5 „π√–À«à“ß∑’Ë¡’°“√‡ª≈’ˬπ√Ÿª ·≈ – ¿“«–∑’Ë¡’§«“¡‡§√’¬ ¥ „π‡™
2010Co-Authors: Sophit Thirach, Nongnuch VanittanakomAbstract:Penicillium marneffei, a Dimorphic Fungus, can cause an opportunistic infection in immunocompromised patients especially in AIDS patients. Molecular genetic studies of P. marneffei provide more understanding about the mechanism of fungal pathogenesis. We have isolated and characterized the ura5 gene, which encodes orotate phosphoribosyltransferase (OPRTase), from P. marneffei. The nucleotide and amino acid sequences of ura5 displayed strong homology to OPRTase proteins in other fungi. Analysis of the ura5 gene expression by RT-PCR revealed that the expression was upregulated at an early step during mycelium to yeast phase transition, and substantially upregulated in mycelial cells exposed to 39oC or treated with 1 mM of H2O2 for 30 min. The results suggested that the ura5 gene might play some role on the thermal adaptation of P. marneffei during temperature-induced yeast phase transition and stress conditions. ∫∑§—¥¬àÕ ‡™◊ÈÕ‡æππ‘´‘‡≈’ˬ¡¡“√å‡πøøî‰Õ®—¥‡ªìπ√ “ Õß√Ÿª∑’Ë “¡“√∂°àÕ„À⇰‘¥°“√µ‘¥‡™◊ÈÕ·∫∫©«¬‚Õ° “ „πºŸâªÉ«¬ ∑’Ë¡’¿Ÿ¡‘§ÿâ¡°—π∫°æ√àÕß‚¥¬‡©æ“–ºŸâªÉ«¬‡Õ ¥ å °“√»÷°…“„π√–¥—∫‚¡‡≈°ÿ≈¢Õ߇™◊ÈÕπ’È®–∑”„Àâ¡’§«“¡‡¢â“„®µàÕ°≈‰° °“√°àÕ‚√§‰¥â¡“°¢÷Èπ „π°“√»÷°…“π’ȉ¥â∑”°“√·¬ ° ·≈–»÷°…“§ÿ≥≈—°…≥–¢Õ߬’π ura5 ‚¥¬¬’ππ’È°”Àπ¥°“ √ √â“ß ‚ª√µ’π orotate phosphoribosyltransferase (OPRTase) º≈°“√»÷°…“æ∫«à “ ≈”¥—∫𑫧≈’‚Õ‰∑¥å·≈–≈”¥—
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Functional Analysis of atfA Gene to Stress Response in Pathogenic Thermal Dimorphic Fungus Penicillium marneffei
2016Co-Authors: Panjaphorn Nimmanee, Patrick C. Y. Woo, Pramote Vanittanakom, Sirida Youngchim, Nongnuch VanittanakomAbstract:Penicillium marneffei, the pathogenic thermal Dimorphic Fungus is a causative agent of a fatal systemic disease, penicilliosis marneffei, in immunocompromised patients especially HIV patients. For growth and survival, this Fungus has to adapt to environmental stresses outside and inside host cells and this adaptation requires stress signaling pathways and regulation of gene expression under various kinds of stresses. In this report, P. marneffei activating transcription factor (atfA) gene encoding bZip-type transcription factor was characterized. To determine functions of this gene, atfA isogenic mutant strain was constructed using the modified split marker recombination method. The phenotypes and susceptibility to varieties of stresses including osmotic, oxidative, heat, UV, cell wall and cell membrane stresses of the mutant strain were compared with the wild type and the atfA complemented strains. Results demonstrated that the mRNA expression level of P. marneffei atfA gene increased under heat stress at 42uC. The atfA mutant was more sensitive to sodium dodecyl sulphate, amphotericin B and tert-butyl hydroperoxide than the wild type and complemented strains but not hydrogen peroxide, menadione, NaCl, sorbitol, calcofluor white, itraconazole, UV stresses and heat stress at 39uC. In addition, recovery of atfA mutant conidia after mouse and human macrophage infections was significantly decreased compared to those of wild type and complemented strains. These results indicated that the atfA gene was required by P. marneffei under specific stres
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characterization of saka gene from pathogenic Dimorphic Fungus penicillium marneffei
International Journal of Medical Microbiology, 2015Co-Authors: Panjaphorn Nimmanee, Patrick C. Y. Woo, Aksarakorn Kummasook, Nongnuch VanittanakomAbstract:Eukaryotes utilize stress activated protein kinase (SAPK) pathways to adapt to environmental stress, including heat, osmotic, oxidative or nutrient stresses. Penicillium marneffei (Talaromyces marneffei), the Dimorphic pathogenic Fungus that can cause disseminated mycosis in HIV-infected patients, has to encounter various types of stresses both outside and inside host cells. However, the strategies used by this Fungus in response to these stresses are still unclear. In this report, the stress-activated kinase (sakA) gene of P. marneffei was characterized and the roles of this gene on various stress conditions were studied. The sakA gene deletion mutant was constructed using the split marker method. The phenotypes and sensitivities to varieties of stresses, including osmotic, oxidative, heat and cell wall stresses of the deletion mutant were compared with the wild type and the sakA complemented strains. Results demonstrated that the P. marneffei sakA gene encoded a putative protein containing TXY phosphorylation lip found in the stress high osmolarity glycerol 1 (Hog1)/Spc1/p38 MAPK family, and that this gene was involved not only in tolerance against oxidative and heat stresses, but also played a role in asexual development, chitin deposition, yeast cell generation in vitro and survival inside mouse and human macrophages.
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functional analysis of atfa gene to stress response in pathogenic thermal Dimorphic Fungus penicillium marneffei
PLOS ONE, 2014Co-Authors: Panjaphorn Nimmanee, Patrick C. Y. Woo, Pramote Vanittanakom, Sirida Youngchim, Nongnuch VanittanakomAbstract:Penicillium marneffei, the pathogenic thermal Dimorphic Fungus is a causative agent of a fatal systemic disease, penicilliosis marneffei, in immunocompromised patients especially HIV patients. For growth and survival, this Fungus has to adapt to environmental stresses outside and inside host cells and this adaptation requires stress signaling pathways and regulation of gene expression under various kinds of stresses. In this report, P. marneffei activating transcription factor (atfA) gene encoding bZip-type transcription factor was characterized. To determine functions of this gene, atfA isogenic mutant strain was constructed using the modified split marker recombination method. The phenotypes and susceptibility to varieties of stresses including osmotic, oxidative, heat, UV, cell wall and cell membrane stresses of the mutant strain were compared with the wild type and the atfA complemented strains. Results demonstrated that the mRNA expression level of P. marneffei atfA gene increased under heat stress at 42°C. The atfA mutant was more sensitive to sodium dodecyl sulphate, amphotericin B and tert-butyl hydroperoxide than the wild type and complemented strains but not hydrogen peroxide, menadione, NaCl, sorbitol, calcofluor white, itraconazole, UV stresses and heat stress at 39°C. In addition, recovery of atfA mutant conidia after mouse and human macrophage infections was significantly decreased compared to those of wild type and complemented strains. These results indicated that the atfA gene was required by P. marneffei under specific stress conditions and might be necessary for fighting against host immune cells during the initiation of infection.