Affiliation:
1. Department of Microbiology, University of Alabama, Birmingham 35294-0005.
Abstract
To examine the cis-acting requirements for RNA replication, a cDNA clone of flock house virus (FHV) RNA 2 was transfected into baby hamster kidney cells and transcribed to yield RNAs that had terminal extensions of different lengths or that lacked internal regions of the molecule. These RNAs were tested for their ability to be replicated by FHV replicase that was provided by cotransfection of purified FHV RNA 1. The results showed that RNA replication was inhibited by terminal extensions, particularly those at the 5' end of the RNA, despite the fact that these extensions were corrected during RNA replication. A negative-sense transcript with a 12-nucleotide 3' extension was replicated to produce a positive-sense RNA that had the correct 5' end, showing that the replicase could select its correct initiation site from within a longer template. A uridine residue at the second position of the positive strand was an important determinant of template activity. RNA molecules with large internal deletions that amounted to almost 50% of the 1,400 nucleotides of RNA 2 replicated as efficiently as full-length molecules, but only if they contained an internal region that lay between nucleotides 538 and 616. Thirty-six spontaneous deletions of RNA 2 that arose during sequential replicative passages all conserved the same internal region of the molecule. These results establish that both terminal and internal regions of FHV RNA 2 play essential roles in making the molecule a competent template for replication.
Publisher
American Society for Microbiology
Subject
Virology,Insect Science,Immunology,Microbiology
Reference29 articles.
1. Cellular expression of a functional nodavirus RNA replicon from vaccinia virus vectors;Ball L. A.;J. Virol.,1992
2. Replication of nodamura virus after transfection of viral RNA into mammalian cells in culture;Ball L. A.;J. Virol.,1992
3. .Ball L. A. and G. W. Wertz. Unpublished data.
4. Ball L. A. B. Wohlrab and Y. Li. Nodavirus RNA replication: mechanism and harnessing to vaccinia virus recombinants. In M. Brinton and R. R. Rueckert (ed.) Molecular biology of positive strand RNA viruses in press.
5. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels;Bonner W. M.;Eur. J. Biochem.,1974
Cited by
70 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献