|
Originally published In Press as doi:10.1074/jbc.M406315200 on July 7, 2004
J. Biol. Chem., Vol. 279, Issue 40, 41384-41392, October 1, 2004
Regulation of Hepatitis C Virus Polyprotein Processing by Signal Peptidase Involves Structural Determinants at the p7 Sequence Junctions*
Séverine Carrère-Kremer ,
Claire Montpellier ,
Lazaro Lorenzo ,
Bénédicte Brulin ,
Laurence Cocquerel ,
Sandrine Belouzard ,
François Penin , and
Jean Dubuisson ¶
From the
CNRS-UPR2511, Institut de Biologie de Lille, 59021 Lille, France and the CNRS-UMR5086, IFR128 BioSciences Lyon-Gerland, Institut de Biologie et de Chimie des Protéines, 69367 Lyon, France
The hepatitis C virus genome encodes a polyprotein precursor that is co- and post-translationally processed by cellular and viral proteases to yield 10 mature protein products (C, E1, E2, p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B). Although most cleavages in hepatitis C virus polyprotein precursor proceed to completion during or immediately after translation, the cleavages mediated by a host cell signal peptidase are partial at the E2/p7 and p7/NS2 sites, leading to the production of an E2p7NS2 precursor. The sequences located immediately N-terminally of E2/p7 and p7/NS2 cleavage sites can function as signal peptides. When fused to a reporter protein, the signal peptides of p7 and NS2 were efficiently cleaved. However, when full-length p7 was fused to the reporter protein, partial cleavage was observed, indicating that a sequence located N-terminally of the signal peptide reduces the efficiency of p7/NS2 cleavage. Sequence analyses and mutagenesis studies have also identified structural determinants responsible for the partial cleavage at both the E2/p7 and p7/NS2 sites. Finally, the short distance between the cleavage site of E2/p7 or p7/NS2 and the predicted transmembrane -helix within the P' region might impose additional structural constraints to the cleavage sites. The insertion of a linker polypeptide sequence between P-3' and P-4' of the cleavage site released these constraints and led to improved cleavage efficiency. Such constraints in the processing of a polyprotein precursor are likely essential for hepatitis C virus to post-translationally regulate the kinetics and/or the level of expression of p7 as well as NS2 and E2 mature proteins.
Received for publication, June 7, 2004
, and in revised form, July 1, 2004.
* This work was supported by the CNRS, European Union Grant QLK21999-00356, a grant from the Association Nationale de Recherche sur le SIDA, and a grant from the Association pour la Recherche sur le Cancer. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
¶ To whom correspondence should be addressed: CNRS-UPR2511, Institut de Biologie de Lille, 1 rue Calmette, BP447, 59021 Lille, France. Tel.: 33-3-20-87-11-60; Fax: 33-3-20-87-12-01; E-mail: jean.dubuisson{at}ibl.fr.

CiteULike Complore Connotea Del.icio.us Digg Reddit Technorati What's this?
This article has been cited by other articles:

|
 |

|
 |
 
V. Jirasko, R. Montserret, N. Appel, A. Janvier, L. Eustachi, C. Brohm, E. Steinmann, T. Pietschmann, F. Penin, and R. Bartenschlager
Structural and Functional Characterization of Nonstructural Protein 2 for Its Role in Hepatitis C Virus Assembly
J. Biol. Chem.,
October 17, 2008;
283(42):
28546 - 28562.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. L. Murray, C. T. Jones, J. Tassello, and C. M. Rice
Alanine Scanning of the Hepatitis C Virus Core Protein Reveals Numerous Residues Essential for Production of Infectious Virus
J. Virol.,
October 1, 2007;
81(19):
10220 - 10231.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
C. T. Jones, C. L. Murray, D. K. Eastman, J. Tassello, and C. M. Rice
Hepatitis C Virus p7 and NS2 Proteins Are Essential for Production of Infectious Virus
J. Virol.,
August 15, 2007;
81(16):
8374 - 8383.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
M. Yi, Y. Ma, J. Yates, and S. M. Lemon
Compensatory Mutations in E1, p7, NS2, and NS3 Enhance Yields of Cell Culture-Infectious Intergenotypic Chimeric Hepatitis C Virus
J. Virol.,
January 15, 2007;
81(2):
629 - 638.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
G. Haqshenas, J. M. Mackenzie, X. Dong, and E. J. Gowans
Hepatitis C virus p7 protein is localized in the endoplasmic reticulum when it is encoded by a replication-competent genome
J. Gen. Virol.,
January 1, 2007;
88(1):
134 - 142.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
T. Pietschmann, A. Kaul, G. Koutsoudakis, A. Shavinskaya, S. Kallis, E. Steinmann, K. Abid, F. Negro, M. Dreux, F.-L. Cosset, et al.
Construction and characterization of infectious intragenotypic and intergenotypic hepatitis C virus chimeras
PNAS,
May 9, 2006;
103(19):
7408 - 7413.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
S. Griffin, D. Clarke, C. McCormick, D. Rowlands, and M. Harris
Signal Peptide Cleavage and Internal Targeting Signals Direct the Hepatitis C Virus p7 Protein to Distinct Intracellular Membranes
J. Virol.,
December 15, 2005;
79(24):
15525 - 15536.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
Y. V. Svitkin, A. Pause, M. Lopez-Lastra, S. Perreault, and N. Sonenberg
Complete Translation of the Hepatitis C Virus Genome In Vitro: Membranes Play a Critical Role in the Maturation of All Virus Proteins except for NS3
J. Virol.,
June 1, 2005;
79(11):
6868 - 6881.
[Abstract]
[Full Text]
[PDF]
|
 |
|

|
 |

|
 |
 
B. J. Isherwood and A. H. Patel
Analysis of the processing and transmembrane topology of the E2p7 protein of hepatitis C virus
J. Gen. Virol.,
March 1, 2005;
86(3):
667 - 676.
[Abstract]
[Full Text]
[PDF]
|
 |
|
Copyright © 2004 by the American Society for Biochemistry and Molecular Biology.
|
Advertisement
Advertisement
|