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Eric L Davis - One of the best experts on this subject based on the ideXlab platform.
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The novel cyst nematode effector protein 19C07 interacts with the Arabidopsis auxin influx transporter LAX3 to control feeding site development.
Plant Physiology, 2011Co-Authors: Chris Lee, Eric L Davis, Thomas J Baum, Demosthenis Chronis, Charlotte Kenning, Benjamin Peret, Tarek Hewezi, Richard Hussey, Malcolm Bennett, Melissa G MitchumAbstract:Plant-parasitic cyst nematodes penetrate plant roots and transform Cells near the vasculature into specialized feeding sites called syncytia. Syncytia form by incorporating neighboring Cells into a single fused Cell by Cell wall dissolution. This process is initiated via injection of esophageal Gland Cell effector proteins from the nematode stylet into the host Cell. Once inside the Cell, these proteins may interact with host proteins that regulate the phytohormone auxin, as Cellular concentrations of auxin increase in developing syncytia. Soybean cyst nematode (Heterodera glycines) Hg19C07 is a novel effector protein expressed specifically in the dorsal Gland Cell during nematode parasitism. Here, we describe its ortholog in the beet cyst nematode (Heterodera schachtii), Hs19C07. We demonstrate that Hs19C07 interacts with the Arabidopsis (Arabidopsis thaliana) auxin influx transporter LAX3. LAX3 is expressed in Cells overlying lateral root primordia, providing auxin signaling that triggers the expression of Cell wall-modifying enzymes, allowing lateral roots to emerge. We found that LAX3 and polygalacturonase, a LAX3-induced Cell wall-modifying enzyme, are expressed in the developing syncytium and in Cells to be incorporated into the syncytium. We observed no decrease in H. schachtii infectivity in aux1 and lax3 single mutants. However, a decrease was observed in both the aux1lax3 double mutant and the aux1lax1lax2lax3 quadruple mutant. In addition, ectopic expression of 19C07 was found to speed up lateral root emergence. We propose that Hs19C07 most likely increases LAX3-mediated auxin influx and may provide a mechanism for cyst nematodes to modulate auxin flow into root Cells, stimulating Cell wall hydrolysis for syncytium development.
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identification of putative parasitism genes expressed in the esophageal Gland Cells of the soybean cyst nematode heterodera glycines
Molecular Plant-microbe Interactions, 2001Co-Authors: Bingli Gao, R Allen, Tom R Maier, Eric L Davis, Thomas J Baum, Richard S HusseyAbstract:Cloning parasitism genes encoding secretory proteins expressed in the esophageal Gland Cells is the key to understanding the molecular basis of nematode parasitism of plants. Suppression subtractive hybridization (SSH) with the microaspirated contents from Heterodera glycines esophageal Gland Cells and intestinal region was used to isolate genes expressed preferentially in the Gland Cells of parasitic stages. Twenty-three unique cDNA sequences from a SSH cDNA library were identified and hybridized to the genomic DNA of H. glycines in Southern blots. Full-length cDNAs of 21 clones were obtained by screening a Gland-Cell long-distance polymerase chain reaction cDNA library. Deduced proteins of 10 clones were preceded by a signal peptide for secretion, and PSORT II computer analysis predicted eight proteins as extraCellular, one as nuclear, and one as plasmalemma localized. In situ hybridization showed that four of the predicted extraCellular clones were expressed specifically in the dorsal Gland Cell, one in the subventral Gland Cells, and three in the intestine in H. glycines. The predicted nuclear clone and the plasmalemma-localized clone were expressed in the subventral Gland Cells and the dorsal Gland Cell, respectively. SSH is an efficient method for cloning putative parasitism genes encoding esophageal Gland Cell secretory proteins that may have a role in H. glycines parasitism of soybean.
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signal peptide selection of cdna cloned directly from the esophageal Gland Cells of the soybean cyst nematode heterodera glycines
Molecular Plant-microbe Interactions, 2001Co-Authors: Xiaohong Wang, R Allen, Tom R Maier, Thomas J Baum, Richard S Hussey, Xiongfei Ding, Melissa Goellner, Jan M De Boer, Eric L DavisAbstract:Secretions from the esophageal Gland Cells of plantparasitic nematodes play critical roles in the nematodeparasitic cycle. A novel method to isolate cDNA encoding putative nematode secretory proteins was developed that utilizes mRNA for reverse transcription-polymerase chain reaction derived from microaspiration of the esophageal Gland Cell contents of parasitic stages of the soybean cyst nematode Heterodera glycines. The resulting H. glycines Gland Cell cDNA was cloned into the pRK18 vector, and plasmid DNA was transformed into a mutated yeast host for specific selection of cDNA inserts that encode proteins with functional signal peptides. Of the 223 cDNA clones recovered from selection in yeast, 97% of the clones encoded a predicted signal peptide. Fourteen unique cDNA clones hybridized to genomic DNA of H. glycines on Southern blots and, among them, nine cDNA clones encoded putative extraCellular proteins, as predicted by PSORT II computer analysis. Four cDNA clones hybridized to transcripts within the...
Hynda K Kleinman - One of the best experts on this subject based on the ideXlab platform.
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role of laminin 1 and tgf beta 3 in acinar differentiation of a human submandibular Gland Cell line hsg
Journal of Cell Science, 1996Co-Authors: Matthew P Hoffman, Maura C Kibbey, John J Letterio, Hynda K KleinmanAbstract:Previous studies show that culturing an immortalized human submandibular Gland Cell line (HSG) on Matrigel, a basement membrane extract, induces cytodifferentiation. We have further defined this model system and identified factors involved in HSG Cell acinar development and cyto-differentiation. Acinar development is marked by Cell migration into multi-Cellular spherical structures, Cell proliferation and apoptosis of the centrally localized Cells. In addition, functional differentiation was determined by indirect immunofluorescence and immunoblot analysis for cystatin, a salivary Gland acinar Cell-specific protein found to be produced by differentiated HSG Cells. Matrigel contains multiple extraCellular matrix proteins, however, laminin-1 was identified as the major matrix component that induced HSG Cell acinar development and cytodifferentiation. Antibodies against specific components of Matrigel and against Cell surface adhesion molecules were added to Cells in culture to identify components important for HSG Cell acinar differentiation. Immunostaining of HSG Cell acini identified TGF-beta 2 and beta 3 as the predominant isoforms within the Cells. Neutralizing antibodies directed against TGF-beta 3 significantly decreased (P
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role of laminin 1 and tgf beta 3 in acinar differentiation of a human submandibular Gland Cell line hsg
Journal of Cell Science, 1996Co-Authors: Matthew P Hoffman, Maura C Kibbey, John J Letterio, Hynda K KleinmanAbstract:Previous studies show that culturing an immortalized human submandibular Gland Cell line (HSG) on Matrigel, a basement membrane extract, induces cytodifferentiation. We have further defined this model system and identified factors involved in HSG Cell acinar development and cyto-differentiation. Acinar development is marked by Cell migration into multi-Cellular spherical structures, Cell proliferation and apoptosis of the centrally localized Cells. In addition, functional differentiation was determined by indirect immunofluorescence and immunoblot analysis for cystatin, a salivary Gland acinar Cell-specific protein found to be produced by differentiated HSG Cells. Matrigel contains multiple extraCellular matrix proteins, however, laminin-1 was identified as the major matrix component that induced HSG Cell acinar development and cytodifferentiation. Antibodies against specific components of Matrigel and against Cell surface adhesion molecules were added to Cells in culture to identify components important for HSG Cell acinar differentiation. Immunostaining of HSG Cell acini identified TGF-beta 2 and beta 3 as the predominant isoforms within the Cells. Neutralizing antibodies directed against TGF-beta 3 significantly decreased (P < or = 0.0002) the size of acini formed. These results indicate that multiple components, including laminin-1 and TGF-beta 3, contribute to HSG Cell acinar development. This model system will be useful to study acinar differentiation and salivary Gland-specific protein expression in vitro.
Thomas J Baum - One of the best experts on this subject based on the ideXlab platform.
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The novel cyst nematode effector protein 19C07 interacts with the Arabidopsis auxin influx transporter LAX3 to control feeding site development.
Plant Physiology, 2011Co-Authors: Chris Lee, Eric L Davis, Thomas J Baum, Demosthenis Chronis, Charlotte Kenning, Benjamin Peret, Tarek Hewezi, Richard Hussey, Malcolm Bennett, Melissa G MitchumAbstract:Plant-parasitic cyst nematodes penetrate plant roots and transform Cells near the vasculature into specialized feeding sites called syncytia. Syncytia form by incorporating neighboring Cells into a single fused Cell by Cell wall dissolution. This process is initiated via injection of esophageal Gland Cell effector proteins from the nematode stylet into the host Cell. Once inside the Cell, these proteins may interact with host proteins that regulate the phytohormone auxin, as Cellular concentrations of auxin increase in developing syncytia. Soybean cyst nematode (Heterodera glycines) Hg19C07 is a novel effector protein expressed specifically in the dorsal Gland Cell during nematode parasitism. Here, we describe its ortholog in the beet cyst nematode (Heterodera schachtii), Hs19C07. We demonstrate that Hs19C07 interacts with the Arabidopsis (Arabidopsis thaliana) auxin influx transporter LAX3. LAX3 is expressed in Cells overlying lateral root primordia, providing auxin signaling that triggers the expression of Cell wall-modifying enzymes, allowing lateral roots to emerge. We found that LAX3 and polygalacturonase, a LAX3-induced Cell wall-modifying enzyme, are expressed in the developing syncytium and in Cells to be incorporated into the syncytium. We observed no decrease in H. schachtii infectivity in aux1 and lax3 single mutants. However, a decrease was observed in both the aux1lax3 double mutant and the aux1lax1lax2lax3 quadruple mutant. In addition, ectopic expression of 19C07 was found to speed up lateral root emergence. We propose that Hs19C07 most likely increases LAX3-mediated auxin influx and may provide a mechanism for cyst nematodes to modulate auxin flow into root Cells, stimulating Cell wall hydrolysis for syncytium development.
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identification of putative parasitism genes expressed in the esophageal Gland Cells of the soybean cyst nematode heterodera glycines
Molecular Plant-microbe Interactions, 2001Co-Authors: Bingli Gao, R Allen, Tom R Maier, Eric L Davis, Thomas J Baum, Richard S HusseyAbstract:Cloning parasitism genes encoding secretory proteins expressed in the esophageal Gland Cells is the key to understanding the molecular basis of nematode parasitism of plants. Suppression subtractive hybridization (SSH) with the microaspirated contents from Heterodera glycines esophageal Gland Cells and intestinal region was used to isolate genes expressed preferentially in the Gland Cells of parasitic stages. Twenty-three unique cDNA sequences from a SSH cDNA library were identified and hybridized to the genomic DNA of H. glycines in Southern blots. Full-length cDNAs of 21 clones were obtained by screening a Gland-Cell long-distance polymerase chain reaction cDNA library. Deduced proteins of 10 clones were preceded by a signal peptide for secretion, and PSORT II computer analysis predicted eight proteins as extraCellular, one as nuclear, and one as plasmalemma localized. In situ hybridization showed that four of the predicted extraCellular clones were expressed specifically in the dorsal Gland Cell, one in the subventral Gland Cells, and three in the intestine in H. glycines. The predicted nuclear clone and the plasmalemma-localized clone were expressed in the subventral Gland Cells and the dorsal Gland Cell, respectively. SSH is an efficient method for cloning putative parasitism genes encoding esophageal Gland Cell secretory proteins that may have a role in H. glycines parasitism of soybean.
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signal peptide selection of cdna cloned directly from the esophageal Gland Cells of the soybean cyst nematode heterodera glycines
Molecular Plant-microbe Interactions, 2001Co-Authors: Xiaohong Wang, R Allen, Tom R Maier, Thomas J Baum, Richard S Hussey, Xiongfei Ding, Melissa Goellner, Jan M De Boer, Eric L DavisAbstract:Secretions from the esophageal Gland Cells of plantparasitic nematodes play critical roles in the nematodeparasitic cycle. A novel method to isolate cDNA encoding putative nematode secretory proteins was developed that utilizes mRNA for reverse transcription-polymerase chain reaction derived from microaspiration of the esophageal Gland Cell contents of parasitic stages of the soybean cyst nematode Heterodera glycines. The resulting H. glycines Gland Cell cDNA was cloned into the pRK18 vector, and plasmid DNA was transformed into a mutated yeast host for specific selection of cDNA inserts that encode proteins with functional signal peptides. Of the 223 cDNA clones recovered from selection in yeast, 97% of the clones encoded a predicted signal peptide. Fourteen unique cDNA clones hybridized to genomic DNA of H. glycines on Southern blots and, among them, nine cDNA clones encoded putative extraCellular proteins, as predicted by PSORT II computer analysis. Four cDNA clones hybridized to transcripts within the...
Richard S Hussey - One of the best experts on this subject based on the ideXlab platform.
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identification of putative parasitism genes expressed in the esophageal Gland Cells of the soybean cyst nematode heterodera glycines
Molecular Plant-microbe Interactions, 2001Co-Authors: Bingli Gao, R Allen, Tom R Maier, Eric L Davis, Thomas J Baum, Richard S HusseyAbstract:Cloning parasitism genes encoding secretory proteins expressed in the esophageal Gland Cells is the key to understanding the molecular basis of nematode parasitism of plants. Suppression subtractive hybridization (SSH) with the microaspirated contents from Heterodera glycines esophageal Gland Cells and intestinal region was used to isolate genes expressed preferentially in the Gland Cells of parasitic stages. Twenty-three unique cDNA sequences from a SSH cDNA library were identified and hybridized to the genomic DNA of H. glycines in Southern blots. Full-length cDNAs of 21 clones were obtained by screening a Gland-Cell long-distance polymerase chain reaction cDNA library. Deduced proteins of 10 clones were preceded by a signal peptide for secretion, and PSORT II computer analysis predicted eight proteins as extraCellular, one as nuclear, and one as plasmalemma localized. In situ hybridization showed that four of the predicted extraCellular clones were expressed specifically in the dorsal Gland Cell, one in the subventral Gland Cells, and three in the intestine in H. glycines. The predicted nuclear clone and the plasmalemma-localized clone were expressed in the subventral Gland Cells and the dorsal Gland Cell, respectively. SSH is an efficient method for cloning putative parasitism genes encoding esophageal Gland Cell secretory proteins that may have a role in H. glycines parasitism of soybean.
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signal peptide selection of cdna cloned directly from the esophageal Gland Cells of the soybean cyst nematode heterodera glycines
Molecular Plant-microbe Interactions, 2001Co-Authors: Xiaohong Wang, R Allen, Tom R Maier, Thomas J Baum, Richard S Hussey, Xiongfei Ding, Melissa Goellner, Jan M De Boer, Eric L DavisAbstract:Secretions from the esophageal Gland Cells of plantparasitic nematodes play critical roles in the nematodeparasitic cycle. A novel method to isolate cDNA encoding putative nematode secretory proteins was developed that utilizes mRNA for reverse transcription-polymerase chain reaction derived from microaspiration of the esophageal Gland Cell contents of parasitic stages of the soybean cyst nematode Heterodera glycines. The resulting H. glycines Gland Cell cDNA was cloned into the pRK18 vector, and plasmid DNA was transformed into a mutated yeast host for specific selection of cDNA inserts that encode proteins with functional signal peptides. Of the 223 cDNA clones recovered from selection in yeast, 97% of the clones encoded a predicted signal peptide. Fourteen unique cDNA clones hybridized to genomic DNA of H. glycines on Southern blots and, among them, nine cDNA clones encoded putative extraCellular proteins, as predicted by PSORT II computer analysis. Four cDNA clones hybridized to transcripts within the...
Matthew P Hoffman - One of the best experts on this subject based on the ideXlab platform.
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role of laminin 1 and tgf beta 3 in acinar differentiation of a human submandibular Gland Cell line hsg
Journal of Cell Science, 1996Co-Authors: Matthew P Hoffman, Maura C Kibbey, John J Letterio, Hynda K KleinmanAbstract:Previous studies show that culturing an immortalized human submandibular Gland Cell line (HSG) on Matrigel, a basement membrane extract, induces cytodifferentiation. We have further defined this model system and identified factors involved in HSG Cell acinar development and cyto-differentiation. Acinar development is marked by Cell migration into multi-Cellular spherical structures, Cell proliferation and apoptosis of the centrally localized Cells. In addition, functional differentiation was determined by indirect immunofluorescence and immunoblot analysis for cystatin, a salivary Gland acinar Cell-specific protein found to be produced by differentiated HSG Cells. Matrigel contains multiple extraCellular matrix proteins, however, laminin-1 was identified as the major matrix component that induced HSG Cell acinar development and cytodifferentiation. Antibodies against specific components of Matrigel and against Cell surface adhesion molecules were added to Cells in culture to identify components important for HSG Cell acinar differentiation. Immunostaining of HSG Cell acini identified TGF-beta 2 and beta 3 as the predominant isoforms within the Cells. Neutralizing antibodies directed against TGF-beta 3 significantly decreased (P
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role of laminin 1 and tgf beta 3 in acinar differentiation of a human submandibular Gland Cell line hsg
Journal of Cell Science, 1996Co-Authors: Matthew P Hoffman, Maura C Kibbey, John J Letterio, Hynda K KleinmanAbstract:Previous studies show that culturing an immortalized human submandibular Gland Cell line (HSG) on Matrigel, a basement membrane extract, induces cytodifferentiation. We have further defined this model system and identified factors involved in HSG Cell acinar development and cyto-differentiation. Acinar development is marked by Cell migration into multi-Cellular spherical structures, Cell proliferation and apoptosis of the centrally localized Cells. In addition, functional differentiation was determined by indirect immunofluorescence and immunoblot analysis for cystatin, a salivary Gland acinar Cell-specific protein found to be produced by differentiated HSG Cells. Matrigel contains multiple extraCellular matrix proteins, however, laminin-1 was identified as the major matrix component that induced HSG Cell acinar development and cytodifferentiation. Antibodies against specific components of Matrigel and against Cell surface adhesion molecules were added to Cells in culture to identify components important for HSG Cell acinar differentiation. Immunostaining of HSG Cell acini identified TGF-beta 2 and beta 3 as the predominant isoforms within the Cells. Neutralizing antibodies directed against TGF-beta 3 significantly decreased (P < or = 0.0002) the size of acini formed. These results indicate that multiple components, including laminin-1 and TGF-beta 3, contribute to HSG Cell acinar development. This model system will be useful to study acinar differentiation and salivary Gland-specific protein expression in vitro.