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. 2007 Jan 20;14(1):31–41. doi: 10.1007/s11373-006-9127-1

Characterization of the cleavage of signal peptide at the C-terminus of hepatitis C virus core protein by signal peptide peptidase

Hsin-Chieh Ma 1, Yi-Yung Ku 2, Yi-Ching Hsieh 2, Shih-Yen Lo 1,2,3,✉
PMCID: PMC7088784  PMID: 17237979

Abstract

Production of hepatitis C virus (HCV) core protein requires the cleavages of polyprotein by signal peptidase and signal peptide peptidase (SPP). Cleavage of signal peptide at the C-terminus of HCV core protein by SPP was characterized in this study. The spko mutant (mutate a.a. 189–193 from ASAYQ to PPFPF) is more efficient than the A/F mutant (mutate a.a 189 and 191 from A to F) in blocking the cleavage of signal peptide by signal peptidase. The cleavage efficiency of SPP is inversely proportional to the length of C-terminal extension of the signal peptide: the longer the extension, the less efficiency the cleavage is. Thus, reducing the length of C-terminal extension of signal peptide by signal peptidase cleavage could facilitate further cleavage by SPP. The recombinant core protein fused with signal peptide from the C-terminus of p7 protein, but not those from the C-termini of E1 and E2, could be cleaved by SPP. Therefore, the sequence of the signal peptide is important but not the sole determinant for its cleavage by SPP. Replacement of the HCV core protein E.R.-associated domain (a.a. 120–150) with the E.R.-associated domain (a.a.1–50) of SARS-CoV membrane protein results in the failure of cleavage of this recombinant protein by SPP, though this protein still is E.R.-associated. This result suggests that not only E.R.-association but also specific protein sequence is important for the HCV core protein signal peptide cleavage by SPP. Thus, our results suggest that both sequences of the signal peptide and the E.R.-associated domain are important for the signal peptide cleavage of HCV core protein by SPP.

Electronic Supplementary MaterialThe online version of this article (doi: 10.1007/s11373-006-9127-1) contains supplementary material, which is available to authorized users.

Keywords: core protein, hepatitis C virus, signal peptidase, signal peptide, signal peptide peptidase

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Acknowledgements

We thank Dr. Charles M. Rice for providing p90/HCVFL-long pU plasmid, Dr. Huichun Li for critical review of this manuscript. This work has been supported by grants from National Science Council of Taiwan (NSC 93-2314-B-320-003 and NSC 953112B320001) to Dr. Shih-Yen Lo.

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