Open AccessImpact of Intra- and Interspecies Variation of Occludin on Its Function as Coreceptor for Authentic Hepatitis C Virus Particles
Author(s) -
Sandra Ciesek,
Sandra Westhaus,
Melanie Wicht,
Ilka Wappler,
Sylvana Henschen,
Christoph Sarrazin,
Nabila Hamdi,
Ahmed Ihab Abdelaziz,
Christian P. Strassburg,
Heiner Wedemeyer,
Michael P. Manns,
Thomas Pietschmann,
Thomas von Hahn
Publication year - 2011
Publication title -
journal of virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.617
H-Index - 292
eISSN - 1070-6321
pISSN - 0022-538X
DOI - 10.1128/jvi.00212-11
Subject(s) - biology , occludin , nonsynonymous substitution , hepatitis c virus , population , viral quasispecies , virology , virus , loss function , gene , genetic variation , cell culture , genetics , coding region , tight junction , phenotype , demography , genome , sociology
Hepatitis C virus (HCV) is characterized by a narrow host range and high interindividual variability in the clinical course of infection. Both of these traits are thought to be largely due to genetic variation between species and between individual hosts. The tight junction component occludin (OCLN) is essential for HCV entry into host cells, and the differences between human and murine OCLN are thought to account in part for the inability of HCV to infect mice and hence preclude their use as a convenient small-animal model. This study assesses the impact of genetic variation in OCLN on cell culture-grown HCV (HCVcc) using a newly generated and characterized OCLNlow subclone of the Huh-7.5 cell line (Huh-7.5 subclone in which endogenous OCLN expression has been downregulated by a short hairpin RNA). We report the frequency of coding nonsynonymous single nucleotide polymorphisms, i.e., polymorphisms resulting in amino acid exchanges, present in the human population and determine their ability to function as HCV (co)receptors. Moreover, we show that murine OCLN can sustain HCVcc entry, albeit with about 5-fold reduced efficiency compared to that of human OCLN. This reduction in efficiency is due solely to two amino acid residues previously identified by others using an HCV pseudoparticle approach. Finally, we use the Huh-7.5/OCLNlow cell line to show that HCV spread between neighboring cells is strictly dependent on OCLN.