Roles for endocytic trafficking and phosphatidylinositol 4-kinase III alpha in hepatitis C virus replication. Export

@article{citeulike:4493965, abstract = {Hepatitis C virus (HCV) reorganizes cellular membranes to establish sites of replication. The required host pathways and the mechanism of cellular membrane reorganization are poorly characterized. Therefore, we interrogated a customized small interfering RNA (siRNA) library that targets 140 host membrane-trafficking genes to identify genes required for both HCV subgenomic replication and infectious virus production. We identified 7 host cofactors of viral replication, including Cdc42 and Rock2 (actin polymerization), EEA1 and Rab5A (early endosomes), Rab7L1, and PI3-kinase C2gamma and PI4-kinase IIIalpha (phospholipid metabolism). Studies of drug inhibitors indicate actin polymerization and phospholipid kinase activity are required for HCV replication. We found extensive co-localization of the HCV replicase markers NS5A and double-stranded RNA with Rab5A and partial co-localization with Rab7L1. PI4K-IIIalpha co-localized with NS5A and double-stranded RNA in addition to being present in detergent-resistant membranes containing NS5A. In a comparison of type II and type III PI4-kinases, PI4Ks were not required for HCV entry, and only PI4K-IIIalpha was required for HCV replication. Although PI4K-IIIalpha siRNAs decreased HCV replication and virus production by almost 100\%, they had no effect on initial HCV RNA translation, suggesting that PI4K-IIIalpha functions at a posttranslational stage. Electron microscopy identified the presence of membranous webs, which are thought to be the site of HCV replication, in HCV-infected cells. Pretreatment with PI4K-IIIalpha siRNAs greatly reduced the accumulation of these membranous web structures in HCV-infected cells. We propose that PI4K-IIIalpha plays an essential role in membrane alterations leading to the formation of HCV replication complexes.}, author = {Berger, Kristi L. and Cooper, Jacob D. and Heaton, Nicholas S. and Yoon, Rosa and Oakland, Todd E. and Jordan, Tristan X. and Mateu, Guaniri and Grakoui, Arash and Randall, Glenn}, citeulike-article-id = {4493965}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0902693106}, citeulike-linkout-1 = {http://www.pnas.org/content/106/18/7577.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/106/18/7577.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/19376974}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=19376974}, day = {5}, doi = {10.1073/pnas.0902693106}, issn = {1091-6490}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {entry, hcv}, month = {May}, number = {18}, pages = {7577--7582}, posted-at = {2009-05-08 16:53:57}, priority = {2}, title = {Roles for endocytic trafficking and phosphatidylinositol 4-kinase III alpha in hepatitis C virus replication.}, url = {http://dx.doi.org/10.1073/pnas.0902693106}, volume = {106}, year = {2009} }

TY - JOUR ID - citeulike:4493965 L3 - citeulike-article-id:4493965 N2 - Hepatitis C virus (HCV) reorganizes cellular membranes to establish sites of replication. The required host pathways and the mechanism of cellular membrane reorganization are poorly characterized. Therefore, we interrogated a customized small interfering RNA (siRNA) library that targets 140 host membrane-trafficking genes to identify genes required for both HCV subgenomic replication and infectious virus production. We identified 7 host cofactors of viral replication, including Cdc42 and Rock2 (actin polymerization), EEA1 and Rab5A (early endosomes), Rab7L1, and PI3-kinase C2gamma and PI4-kinase IIIalpha (phospholipid metabolism). Studies of drug inhibitors indicate actin polymerization and phospholipid kinase activity are required for HCV replication. We found extensive co-localization of the HCV replicase markers NS5A and double-stranded RNA with Rab5A and partial co-localization with Rab7L1. PI4K-IIIalpha co-localized with NS5A and double-stranded RNA in addition to being present in detergent-resistant membranes containing NS5A. In a comparison of type II and type III PI4-kinases, PI4Ks were not required for HCV entry, and only PI4K-IIIalpha was required for HCV replication. Although PI4K-IIIalpha siRNAs decreased HCV replication and virus production by almost 100%, they had no effect on initial HCV RNA translation, suggesting that PI4K-IIIalpha functions at a posttranslational stage. Electron microscopy identified the presence of membranous webs, which are thought to be the site of HCV replication, in HCV-infected cells. Pretreatment with PI4K-IIIalpha siRNAs greatly reduced the accumulation of these membranous web structures in HCV-infected cells. We propose that PI4K-IIIalpha plays an essential role in membrane alterations leading to the formation of HCV replication complexes. IS - 18 JF - Proceedings of the National Academy of Sciences of the United States of America SN - 1091-6490 EP - 7582 TI - Roles for endocytic trafficking and phosphatidylinositol 4-kinase III alpha in hepatitis C virus replication. VL - 106 SP - 7577 KW - entry KW - hcv AU - Berger, Kristi AU - Cooper, Jacob AU - Heaton, Nicholas AU - Yoon, Rosa AU - Oakland, Todd AU - Jordan, Tristan AU - Mateu, Guaniri AU - Grakoui, Arash AU - Randall, Glenn PY - 2009/05/05/ UR - http://dx.doi.org/10.1073/pnas.0902693106 ER -