Hepatitis A virus capsid protein VP1 has a heterogeneous C terminus

被引:36
作者
Graff, J
Richards, OC
Swiderek, KM
Davis, MT
Rusnak, F
Harmon, SA
Jia, XY
Summers, DF
Ehrenfeld, E
机构
[1] Univ Calif Irvine, Dept Mol Biol & Biochem, Irvine, CA 92697 USA
[2] Univ Calif Irvine, Dept Microbiol & Mol Genet, Irvine, CA 92697 USA
[3] Inst City Hope, Beckman Res Inst, Duarte, CA 91010 USA
关键词
D O I
10.1128/JVI.73.7.6015-6023.1999
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Hepatitis A virus (HAV) encodes a single polyprotein which is posttranslationally processed into the functional structural and nonstructural proteins. Only one protease, viral protease 3C, has been implicated in the nine protein scissions. Processing of the capsid protein precursor region generates a unique intermediate, PX (VP1-2A), which accumulates in infected cells and is assumed to serve as precursor to VP1 found in virions, although the details of this reaction have not been determined. Coexpression in transfected cells of a variety of P1 precursor proteins with viral protease 3C demonstrated efficient production of PX, as well as VP0 and VP3; however, no mature VP1 protein was detected. To identify the C-terminal amino acid residue of HAV VPI, we performed peptide sequence analysis by protease-catalyzed [O-18]H2O incorporation followed by liquid chromatography ion-trap microspray tandem mass spectrometry of HAV VP1 isolated from purified virions. Two different cell culture-adapted isolates of HAV, strains HM175pE and HM175p35, were used for these analyses. VP1 preparations from both virus isolates contained heterogeneous C termini. The predominant C-terminal amino acid in both virus preparations was VP1-Ser274, which is located N terminal to a methionine residue in VP1-2A. In addition, the analysis of HM175pE recovered smaller amounts of amino acids VP1-Glu273 and VP1-Thr272. In the case of HM175p35, which contains valine at amino acid position VP1-273, VP1-Thr272 was found in addition to VP1-Ser274. The data suggest that HAV 3C is not the protease responsible for generation of the VP1 C terminus. We propose the involvement of host cell protease(s) in the production of HAV VP1.
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页码:6015 / 6023
页数:9
相关论文
共 47 条
[1]   MORPHOGENESIS OF HEPATITIS-A VIRUS - ISOLATION AND CHARACTERIZATION OF SUBVIRAL PARTICLES [J].
ANDERSON, DA ;
ROSS, BC .
JOURNAL OF VIROLOGY, 1990, 64 (11) :5284-5289
[2]  
BAILEY JM, 1992, PROTEIN SCI, V1, P68
[3]   The refined crystal structure of the 3C gene product from hepatitis A virus: Specific proteinase activity and RNA recognition [J].
Bergmann, EM ;
Mosimann, SC ;
Chernaia, MM ;
Malcolm, BA ;
James, MNG .
JOURNAL OF VIROLOGY, 1997, 71 (03) :2436-2448
[4]   Hepatitis A virus subviral particles: purification, accumulation, and relative infectivity of virions, provirions and procapsids [J].
Bishop, NE ;
Anderson, DA .
ARCHIVES OF VIROLOGY, 1997, 142 (11) :2147-2160
[5]   MENGO-VIRUS MATURATION IS ACCOMPANIED BY C-TERMINAL MODIFICATION OF CAPSID PROTEIN-VP1 [J].
BOEGE, U ;
SCRABA, DG .
VIROLOGY, 1989, 168 (02) :409-412
[6]   SYNTHESIS AND ASSEMBLY OF HEPATITIS-A VIRUS-SPECIFIC PROTEINS IN BS-C-1 CELLS [J].
BOROVEC, SV ;
ANDERSON, DA .
JOURNAL OF VIROLOGY, 1993, 67 (06) :3095-3102
[7]   RAPID COMPLETION OF THE REPLICATION CYCLE OF HEPATITIS-A VIRUS SUBSEQUENT TO REVERSAL OF GUANIDINE INHIBITION [J].
CHO, MW ;
EHRENFELD, E .
VIROLOGY, 1991, 180 (02) :770-780
[8]   HEPATITIS-A VIRUS CDNA AND ITS RNA TRANSCRIPTS ARE INFECTIOUS IN CELL-CULTURE [J].
COHEN, JI ;
TICEHURST, JR ;
FEINSTONE, SM ;
ROSENBLUM, B ;
PURCELL, RH .
JOURNAL OF VIROLOGY, 1987, 61 (10) :3035-3039
[9]   COMPLETE NUCLEOTIDE-SEQUENCE OF WILD-TYPE HEPATITIS-A VIRUS - COMPARISON WITH DIFFERENT STRAINS OF HEPATITIS-A VIRUS AND OTHER PICORNAVIRUSES [J].
COHEN, JI ;
TICEHURST, JR ;
PURCELL, RH ;
BUCKLERWHITE, A ;
BAROUDY, BM .
JOURNAL OF VIROLOGY, 1987, 61 (01) :50-59
[10]   Variable flow liquid chromatography tandem mass spectrometry and the comprehensive analysis of complex protein digest mixtures [J].
Davis, MT ;
Lee, TD .
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 1997, 8 (10) :1059-1069