The second extracellular loop dictates Occludin-mediated HCV entry.
Recent findings have implicated tight junction (TJ) protein Occludin (OCLN) as an essential factor for hepatitis C virus (HCV) to enter human hepatocytes. To gain insights into OCLN-mediated HCV entry, we created a panel of OCLN deletion mutants and found that without impairing OCLN's cell surface localization, removal of the extracellular loop 2 (EL2) from OCLN abolished both its ability to mediate HIV-HCV pseudotypes' (HCVpp) entry as well as its ability to coprecipitate HCV glycoprotein E2. Recombinant OCLN EL2, however, failed to robustly bind soluble E2 (sE2) in pull-down assays. Subsequent studies revealed that OCLN formed complex with Dynamin II, an important GTPase for endocytosis, in an EL2-dependent fashion. HCVpp, as well as cell culture grown HCV (HCVcc), was sensitive to Dynamin knockdown or inhibition. We conclude that OCLN EL2 dictates the Dynamin-dependent HCV entry. Furthermore, OCLN could function to bridge virions to Dynamin-dependent endocytic machineries.
Liu
Shufeng
S
Department of Infectious Diseases and Microbiology, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Kuo
Wayne
W
Yang
Wei
W
Liu
Weiqun
W
Gibson
Gregory A
GA
Dorko
Kenneth
K
Watkins
Simon C
SC
Strom
Stephen C
SC
Wang
Tianyi
T
D002460:Cell Line; D034285:Dynamin II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006678:HIV / Q000235:genetics; D016174:Hepacivirus / Q000502:physiology*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D050505:Mutant Proteins / Q000235:genetics / Q000502:physiology; D062793:Occludin; D011485:Protein Binding; D011991:Receptors, Virus / Q000235:genetics / Q000502:physiology*; D017384:Sequence Deletion; D053586:Virus Internalization
D008565:Membrane Proteins; D050505:Mutant Proteins; C568985:OCLN protein, human; D062793:Occludin; D011991:Receptors, Virus; D034285:Dynamin II
J Virol. 2000 Jan;74(2):702-9
10623732
Hepatology. 2008 Oct;48(4):1044-53
18802961
J Virol. 1999 Aug;73(8):6235-44
10400713
J Virol. 2008 Sep;82(17):8316-29
18579606
J Virol. 2010 Jul;84(14):6987-94
20463075
Nature. 2007 Apr 12;446(7137):801-5
17325668
Oncogene. 2008 Nov 24;27(55):6930-8
19029935
EMBO J. 2007 Sep 5;26(17):4016-28
17717529
Virol J. 2008 Mar 20;5:46
18355410
J Virol. 2008 Jan;82(1):461-70
17959672
Biochim Biophys Acta. 2009 Apr;1788(4):864-71
18952050
Drug Metab Dispos. 2004 May;32(5):512-8
15100173
FASEB J. 2005 Jun;19(8):923-33
15923402
J Virol. 1999 Apr;73(4):2641-9
10074109
J Cell Sci. 1997 May;110 ( Pt 9):1113-21
9175707
J Cell Sci. 1998 Feb;111 ( Pt 4):511-9
9443899
Mol Biol Cell. 2000 Dec;11(12):4131-42
11102513
J Cell Sci. 1999 Jun;112 ( Pt 12):1879-88
10341207
Mol Cell Proteomics. 2007 Nov;6(11):1855-67
17644758
Mol Biol Cell. 2004 Jan;15(1):176-88
14528017
Hepatology. 2008 Nov;48(5):1396-403
18830996
J Virol. 2009 Aug;83(16):8012-20
19515778
Proc Natl Acad Sci U S A. 2009 Dec 8;106(49):20770-5
19948954
Nature. 2009 Feb 12;457(7231):882-6
19182773
Adv Drug Deliv Rev. 2005 Apr 25;57(6):883-917
15820558
EMBO J. 2002 Oct 1;21(19):5017-25
12356718
Methods Enzymol. 1996;272:388-401
8791798
PLoS Pathog. 2009 Apr;5(4):e1000394
19390604
FEBS Lett. 2000 Jul 7;476(3):258-61
10913624
Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):7271-6
12761383
Biochim Biophys Acta. 2005 May 15;1669(1):34-42
15842997
J Biol Chem. 2008 Mar 28;283(13):8643-53
18211898
J Biol Chem. 2010 Jul 2;285(27):21092-102
20375010
J Virol. 2008 May;82(10):5007-20
18337570
J Virol. 2006 Jul;80(14):6964-72
16809302
J Gen Virol. 2006 Sep;87(Pt 9):2583-2593
16894197
Oncogene. 2005 Jun 23;24(27):4412-20
15806147
J Exp Med. 2003 Mar 3;197(5):633-42
12615904
Nat Rev Mol Cell Biol. 2004 Feb;5(2):133-47
15040446
Virology. 2006 Apr 25;348(1):1-12
16455127
J Virol. 2003 Oct;77(19):10677-83
12970454
Proc Natl Acad Sci U S A. 2005 Jun 28;102(26):9294-9
15939869
J Cell Biol. 1997 Jan 27;136(2):399-409
9015310
J Cell Sci. 2008 Nov 15;121(Pt 22):3677-82
18987354
J Endocrinol. 2006 Dec;191(3):571-86
17170215
Cell Host Microbe. 2007 Sep 13;2(3):181-92
18005733
Science. 1998 Oct 30;282(5390):938-41
9794763
J Virol. 2009 Feb;83(4):2011-4
19052094
J Virol. 2006 Dec;80(23):11571-8
17005647
J Cell Biol. 1993 Dec;123(6 Pt 2):1777-88
8276896