Identification of a specific interaction between the coronavirus mouse hepatitis virus A59 nucleocapsid protein and packaging signal

被引:55
作者
Molenkamp, R [1 ]
Spaan, WJM [1 ]
机构
[1] Leiden Univ, Inst Med Microbiol, Dept Virol, NL-2300 RC Leiden, Netherlands
关键词
D O I
10.1006/viro.1997.8867
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The coronavirus mouse hepatitis virus (MHV) is an enveloped positive stranded RNA virus. In infected cells MHV produces a 3' coterminal nested se; of subgenomic messenger RNAs. Only the genomic RNA, however, is encapsidated by the nucleocapsid protein and incorporated in infectious MHV virions. II is believed that an RNA packaging signal (Ps), present only in the genomic RNA, is responsible for this selectivity. Earlier studies mapped this signal to a 69-nt stem-loop structure positioned in the 3' end of ORF1b., The selective encapsidation mechanism probably initiates by specific interaction of the packaging signal with the nucleocapsid protein. In this study we demonstrate the in vitro interaction of the MHV-A59 nucleocapsid protein with the packaging signal of: MHV using gel retardation and UV cross-linking assays. This interaction was observed not only with the nucleocapsid protein from infected cells but also with that from purified virions and from cells expressing a recombinant nucleocapsid protein. The specificity of the interaction was demonstrated by competition experiments with nonlabeled Ps containing RNAs, tRNA, and total cytoplasmic RNA. The results indicated that no virus specific modification of the N-protein or the presence of other viral proteins are required for this in vitro interaction. The assays described in this report provide us With a powerful tool for studying encapsidation (initiation) in more detail. (C) 1997 Academic Press.
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页码:78 / 86
页数:9
相关论文
共 55 条
[1]   MUTATIONS OF RNA AND PROTEIN SEQUENCES INVOLVED IN HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PACKAGING RESULT IN PRODUCTION OF NONINFECTIOUS VIRUS [J].
ALDOVINI, A ;
YOUNG, RA .
JOURNAL OF VIROLOGY, 1990, 64 (05) :1920-1926
[2]   Specificity of ribonucleoprotein interaction determined by RNA folding during complex formation [J].
Allain, FHT ;
Gubser, CC ;
Howe, PWA ;
Nagai, K ;
Neuhaus, D ;
Varani, G .
NATURE, 1996, 380 (6575) :646-650
[3]   MEMBRANE AND PHOSPHOLIPID BINDING BY MURINE CORONAVIRAL NUCLEOCAPSID N-PROTEIN [J].
ANDERSON, R ;
WONG, F .
VIROLOGY, 1993, 194 (01) :224-232
[4]   SEQUENCE OF THE NUCLEOCAPSID GENE FROM MURINE CORONAVIRUS MHV-A59 [J].
ARMSTRONG, J ;
SMEEKENS, S ;
ROTTIER, P .
NUCLEIC ACIDS RESEARCH, 1983, 11 (03) :883-891
[5]   INTERACTIONS BETWEEN CORONAVIRUS NUCLEOCAPSID PROTEIN AND VIRAL RNAS - IMPLICATIONS FOR VIRAL TRANSCRIPTION [J].
BARIC, RS ;
NELSON, GW ;
FLEMING, JO ;
DEANS, RJ ;
KECK, JG ;
CASTEEL, N ;
STOHLMAN, SA .
JOURNAL OF VIROLOGY, 1988, 62 (11) :4280-4287
[6]   Sequence- and structure-specific determinants in the interaction between the RNA encapsidation signal and reverse transcriptase of avian hepatitis B viruses [J].
Beck, J ;
Nassal, M .
JOURNAL OF VIROLOGY, 1997, 71 (07) :4971-4980
[7]   RETROVIRAL NUCLEOCAPSID DOMAINS MEDIATE THE SPECIFIC RECOGNITION OF GENOMIC VIRAL RNAS BY CHIMERIC GAG POLYPROTEINS DURING RNA PACKAGING IN-VIVO [J].
BERKOWITZ, RD ;
OHAGEN, A ;
HOGLUND, S ;
GOFF, SP .
JOURNAL OF VIROLOGY, 1995, 69 (10) :6445-6456
[8]   A subgenomic mRNA transcript of the coronavirus mouse hepatitis virus strain A59 defective interfering (DI) RNA is packaged when it contains the DI packaging signal [J].
Bos, ECW ;
Dobbe, JC ;
Luytjes, W ;
Spaan, WJM .
JOURNAL OF VIROLOGY, 1997, 71 (07) :5684-5687
[9]   The production of recombinant infectious DI-particles of a murine coronavirus in the absence of helper virus [J].
Bos, ECW ;
Luytjes, W ;
VanderMeulen, H ;
Koerten, HK ;
Spaan, WJM .
VIROLOGY, 1996, 218 (01) :52-60
[10]  
BREDENBEEK PJ, 1990, THESIS U UTRECHT UTR