LOCALIZATION OF AN RNA-BINDING DOMAIN IN THE NUCLEOCAPSID PROTEIN OF THE CORONAVIRUS MOUSE HEPATITIS-VIRUS

被引:62
作者
MASTERS, PS [1 ]
机构
[1] SUNY ALBANY,SCH PUBL HLTH,DEPT BIOMED SCI,ALBANY,NY 12222
关键词
D O I
10.1007/BF01309634
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The interaction between the nucleocapsid (N) protein of mouse hepatitis virus (MHV) and RNA was studied in an effort to define portions of the N molecule that participate in binding to RNA. N mRNAs transcribed from SP6 and T7 vectors were translated in a rabbit reticulocyte lysate. Analysis of synthesized N protein in a nondenaturing gel system showed that it bound in vitro to an endogenous RNA in the reticulocyte lysate but not to its own mRNA. A set of deletion mutants was constructed in order to localize the RNA-binding activity of the N protein. It was found that removal of as much as 135 amino-terminal or 57 carboxy-terminal amino acids from the molecule had little or no effect on RNA binding. Moreover, deletion mutants lacking both termini still retained RNA-binding ability. By contrast, internal deletions or truncations extending beyond these two limits effectively abolished RNA binding by N protein. Thus, the RNA-binding region of N has been mapped to the second (central) of the three structural domains of the molecule.
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页码:141 / 160
页数:20
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