IDENTIFICATION, EXPRESSION, AND PROCESSING OF AN 87-KDA POLYPEPTIDE ENCODED BY ORF 1A OF THE CORONAVIRUS INFECTIOUS-BRONCHITIS VIRUS

被引:31
作者
LIU, DX [1 ]
TIBBLES, KW [1 ]
CAVANAGH, D [1 ]
BROWN, TDK [1 ]
BRIERLEY, I [1 ]
机构
[1] AFRC,INST ANIM HLTH,COMPTON LAB,NEWBURY RG16 0NN,BERKS,ENGLAND
基金
英国医学研究理事会;
关键词
D O I
10.1006/viro.1995.1128
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Nucleotide sequence analysis has shown previously that the genomic-length mRNA (mRNA 1) of the coronavirus infectious bronchitis virus (IBV) contains two large open reading frames (ORFs), 1a and 1b, with the potential to encode polyproteins of approximately 441 and 300 kDa, respectively. We have characterized the specificity of a set of region-specific antisera raised against the 5'-portion of ORF la by immunoprecipitation of in vitro-synthesized, C-terminally truncated la polypeptides and used these antisera to detect virus-specific proteins in IBV-infected vero cells. Two antisera, which had specificity for IBV sequences from nucleotides 710 to 2079 and 1355 to 2433, respectively, immunoprecipitated a polypeptide of approximately 87 kDa from IBV-infected Vero cells. In vitro translation of ORF la sequence terminating at nucleotide 5763 did not produce this protein unless the in vitro translation products were incubated with Vero cell sin extracts prepared from either IBV-infected or mock-infected Vero cells, However, processing of the 87-kDa protein was also observed when the same region was expressed in Vero cells using the vaccinia virus/T7 expression system. This observation indicates that the 87-kDa polypeptide is encoded within the 5'-most 3000 nucleotides of mRNA I and that it might be cleaved from the la polyprotein by viral and cellular proteinases. (C) 1995 Academic Press, Inc.
引用
收藏
页码:48 / 57
页数:10
相关论文
共 30 条
[1]   IDENTIFICATION OF A DOMAIN REQUIRED FOR AUTOPROTEOLYTIC CLEAVAGE OF MURINE CORONAVIRUS GENE-A POLYPROTEIN [J].
BAKER, SC ;
SHIEH, CK ;
SOE, LH ;
CHANG, MF ;
VANNIER, DM ;
LAI, MMC .
JOURNAL OF VIROLOGY, 1989, 63 (09) :3693-3699
[2]   IDENTIFICATION OF THE CATALYTIC SITES OF A PAPAIN-LIKE CYSTEINE PROTEINASE OF MURINE CORONAVIRUS [J].
BAKER, SC ;
YOKOMORI, K ;
DONG, S ;
CARLISLE, R ;
GORBALENYA, AE ;
KOONIN, EV ;
LAI, MMC .
JOURNAL OF VIROLOGY, 1993, 67 (10) :6056-6063
[3]  
BLAIR WS, 1993, J VIROL, V67, P2339
[4]   COMPLETION OF THE SEQUENCE OF THE GENOME OF THE CORONAVIRUS AVIAN INFECTIOUS-BRONCHITIS VIRUS [J].
BOURSNELL, MEG ;
BROWN, TDK ;
FOULDS, IJ ;
GREEN, PF ;
TOMLEY, FM ;
BINNS, MM .
JOURNAL OF GENERAL VIROLOGY, 1987, 68 :57-77
[5]  
BRIERLEY I, 1990, ADV EXP MED BIOL, V276, P275
[6]   AN EFFICIENT RIBOSOMAL FRAME-SHIFTING SIGNAL IN THE POLYMERASE-ENCODING REGION OF THE CORONAVIRUS IBV [J].
BRIERLEY, I ;
BOURSNELL, MEG ;
BINNS, MM ;
BILIMORIA, B ;
BLOK, VC ;
BROWN, TDK ;
INGLIS, SC .
EMBO JOURNAL, 1987, 6 (12) :3779-3785
[7]   CHARACTERIZATION OF AN EFFICIENT CORONAVIRUS RIBOSOMAL FRAMESHIFTING SIGNAL - REQUIREMENT FOR AN RNA PSEUDOKNOT [J].
BRIERLEY, I ;
DIGARD, P ;
INGLIS, SC .
CELL, 1989, 57 (04) :537-547
[8]   RECOMMENDATIONS OF THE CORONAVIRUS STUDY-GROUP FOR THE NOMENCLATURE OF THE STRUCTURAL PROTEINS, MESSENGER-RNAS, AND GENES OF CORONAVIRUSES [J].
CAVANAGH, D ;
BRIAN, DA ;
ENJUANES, L ;
HOLMES, KV ;
LAI, MMC ;
LAUDE, H ;
SIDDELL, SG ;
SPAAN, W ;
TAGUCHI, F ;
TALBOT, PJ .
VIROLOGY, 1990, 176 (01) :306-307
[9]   FLAVIVIRUS GENOME ORGANIZATION, EXPRESSION, AND REPLICATION [J].
CHAMBERS, TJ ;
HAHN, CS ;
GALLER, R ;
RICE, CM .
ANNUAL REVIEW OF MICROBIOLOGY, 1990, 44 :649-688
[10]   SIMPLE, EFFICIENT INVITRO SYNTHESIS OF CAPPED RNA USEFUL FOR DIRECT EXPRESSION OF CLONED EUKARYOTIC GENES [J].
CONTRERAS, R ;
CHEROUTRE, H ;
DEGRAVE, W ;
FIERS, W .
NUCLEIC ACIDS RESEARCH, 1982, 10 (20) :6353-6362