The Experts below are selected from a list of 42 Experts worldwide ranked by ideXlab platform

Mark Harris - One of the best experts on this subject based on the ideXlab platform.

  • release of Infectious Hepatitis c virus from huh7 cells occurs via a trans golgi network to endosome pathway independent of very low density lipoprotein secretion
    Journal of Virology, 2016
    Co-Authors: Jamel Mankouri, Cheryl T Walter, Hazel Stewart, Matthew Bentham, Wei Sun Park, Mitsunori Fukuda, Stephen Griffin, Mark Harris
    Abstract:

    ABSTRACT The release of Infectious Hepatitis C virus (HCV) particles from infected cells remains poorly characterized. We previously demonstrated that virus release is dependent on the endosomal sorting complex required for transport (ESCRT). Here, we show a critical role of trans -Golgi network (TGN)-endosome trafficking during the assembly, but principally the secretion, of Infectious virus. This was demonstrated by both small interfering RNA (siRNA)-mediated silencing of TGN-associated adaptor proteins and a panel of dominant negative (DN) Rab GTPases involved in TGN-endosome trafficking steps. Importantly, interfering with factors critical for HCV release did not have a concomitant effect on secretion of triglycerides, ApoB, or ApoE, indicating that particles are likely released from Huh7 cells via pathways distinct from that of very-low-density lipoprotein (VLDL). Finally, we show that HCV NS2 perturbs TGN architecture, redistributing TGN membranes to closely associate with HCV core protein residing on lipid droplets. These findings support the notion that HCV hijacks TGN-endosome trafficking to facilitate particle assembly and release. Moreover, although essential for assembly and infectivity, the trafficking of mature virions is seemingly independent of host lipoproteins. IMPORTANCE The mechanisms by which Infectious Hepatitis C virus particles are assembled and released from the cell are poorly understood. We show that the virus subverts host cell trafficking pathways to effect the release of virus particles and disrupts the structure of the Golgi apparatus, a key cellular organelle involved in secretion. In addition, we demonstrate that the mechanisms used by the virus to exit the cell are distinct from those used by the cell to release lipoproteins, suggesting that the virus effects a unique modification to cellular trafficking pathways.

Bertrand Saunier - One of the best experts on this subject based on the ideXlab platform.

Jamel Mankouri - One of the best experts on this subject based on the ideXlab platform.

  • release of Infectious Hepatitis c virus from huh7 cells occurs via a trans golgi network to endosome pathway independent of very low density lipoprotein secretion
    Journal of Virology, 2016
    Co-Authors: Jamel Mankouri, Cheryl T Walter, Hazel Stewart, Matthew Bentham, Wei Sun Park, Mitsunori Fukuda, Stephen Griffin, Mark Harris
    Abstract:

    ABSTRACT The release of Infectious Hepatitis C virus (HCV) particles from infected cells remains poorly characterized. We previously demonstrated that virus release is dependent on the endosomal sorting complex required for transport (ESCRT). Here, we show a critical role of trans -Golgi network (TGN)-endosome trafficking during the assembly, but principally the secretion, of Infectious virus. This was demonstrated by both small interfering RNA (siRNA)-mediated silencing of TGN-associated adaptor proteins and a panel of dominant negative (DN) Rab GTPases involved in TGN-endosome trafficking steps. Importantly, interfering with factors critical for HCV release did not have a concomitant effect on secretion of triglycerides, ApoB, or ApoE, indicating that particles are likely released from Huh7 cells via pathways distinct from that of very-low-density lipoprotein (VLDL). Finally, we show that HCV NS2 perturbs TGN architecture, redistributing TGN membranes to closely associate with HCV core protein residing on lipid droplets. These findings support the notion that HCV hijacks TGN-endosome trafficking to facilitate particle assembly and release. Moreover, although essential for assembly and infectivity, the trafficking of mature virions is seemingly independent of host lipoproteins. IMPORTANCE The mechanisms by which Infectious Hepatitis C virus particles are assembled and released from the cell are poorly understood. We show that the virus subverts host cell trafficking pathways to effect the release of virus particles and disrupts the structure of the Golgi apparatus, a key cellular organelle involved in secretion. In addition, we demonstrate that the mechanisms used by the virus to exit the cell are distinct from those used by the cell to release lipoproteins, suggesting that the virus effects a unique modification to cellular trafficking pathways.

Miriam Triyatni - One of the best experts on this subject based on the ideXlab platform.

Rosa Aznar - One of the best experts on this subject based on the ideXlab platform.

  • Discrimination of Infectious Hepatitis A Viruses by Propidium Monoazide Real-Time RT-PCR
    Food and Environmental Virology, 2012
    Co-Authors: Gloria Sánchez, Patricia Elizaquivel, Rosa Aznar
    Abstract:

    The discrimination of Infectious and inactivated viruses remains a key obstacle when using quantitative RT-PCR (RT-qPCR) to quantify enteric viruses. In this study, propidium monoazide (PMA) and RNase pretreatments were evaluated for the detection and quantification of Infectious Hepatitis A virus (HAV). For thermally inactivated HAV, PMA treatment was more effective than RNase treatment for differentiating Infectious and inactivated viruses, with HAV titers reduced by more than 2.4 log_10 units. Results showed that combining 50 μM of PMA and RT-qPCR selectively quantify Infectious HAV in media suspensions. Therefore, PMA treatment previous to RT-qPCR detection is a promising alternative to assess HAV infectivity.