SUMMARY: The p7 protein of hepatitis C virus (HCV) has been classified into a family of viral proteins, designated viroporins that form ion channels. The M2 protein of influenza virus is the prototype viroporin and encodes a HXXXW motif that constitutes the main functional element of the M2 channels. Alignment of different p7 proteins revealed that a HXXXW sequence (positions 17-21) is also highly conserved among some HCV genotypes. To study the putative HXXXW motif in p7, five mutants of the Japanese fulminant hepatitis 1 strain of HCV that encoded H17A, H17G, H17E, Y21A and Y21W were generated. After transfection of human hepatoma cells with the mutant transcripts, unlike H17A and H17G that produced up to 1 log lower viral titres than wild type, H17E and Y21W showed slightly higher infectivity. In conclusion, this study demonstrated that the HXXXW sequence exists in the p7 proteins of some HCV genotypes and that H17 plays an important role in virus replication.