@article{TEXTUAL,
      recid = {10529},
      author = {Coller, Kelly E. and Berger, Kristi L. and Heaton,  Nicholas S. and Cooper, Jacob D. and Yoon, Rosa and  Randall, Glenn},
      title = {RNA Interference and Single Particle Tracking Analysis of  Hepatitis C Virus Endocytosis},
      journal = {PLOS Pathology},
      address = {2009-12-24},
      number = {TEXTUAL},
      abstract = {<p>Hepatitis C virus (HCV) enters hepatocytes following a  complex set of receptor interactions, culminating in  internalization via clathrin-mediated endocytosis. However,  aside from receptors, little is known about the cellular  molecular requirements for infectious HCV entry. Therefore,  we analyzed a siRNA library that targets 140 cellular  membrane trafficking genes to identify host genes required  for infectious HCV production and HCV pseudoparticle entry.  This approach identified 16 host cofactors of HCV entry  that function primarily in clathrin-mediated endocytosis,  including components of the clathrin endocytosis machinery,  actin polymerization, receptor internalization and sorting,  and endosomal acidification. We next developed single  particle tracking analysis of highly infectious fluorescent  HCV particles to examine the co-trafficking of HCV virions  with cellular cofactors of endocytosis. We observe  multiple, sequential interactions of HCV virions with the  actin cytoskeleton, including retraction along filopodia,  actin nucleation during internalization, and migration of  internalized particles along actin stress fibers. HCV  co-localizes with clathrin and the ubiquitin ligase c-Cbl  prior to internalization. Entering HCV particles are  associated with the receptor molecules CD81 and the tight  junction protein, claudin-1; however, HCV-claudin-1  interactions were not restricted to Huh-7.5 cell-cell  junctions. Surprisingly, HCV internalization generally  occurred outside of Huh-7.5 cell-cell junctions, which may  reflect the poorly polarized nature of current HCV cell  culture models. Following internalization, HCV particles  transport with GFP-Rab5a positive endosomes, which is  consistent with trafficking to the early endosome. This  study presents technical advances for imaging HCV entry, in  addition to identifying new host cofactors of HCV  infection, some of which may be antiviral targets.</p>},
      url = {http://knowledge.uchicago.edu/record/10529},
}