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Published May 15, 1978 | public
Journal Article

Replication of sindbis virus VI. Poly(A) and poly(U) in virus-specific RNA species

Abstract

Polyadenylic acid [poly(A)] isolated from all species of Sindbis virus RNA is heterogeneous in size with a mean length of 70 nucleotides and a size range of from 46 to 296 nucleotides. Poly(A)s isolated from Sindbis virion RNA grown in chicken, hamster, and mosquito culture cells have similar size distributions. Base composition analysis of Sindbis poly(A) indicates that it is preceded on its 5'-end by a short stretch of pyrimidine residues. Oligo(dT)-cellulose chromatography of whole viral RNA reveals that most Sindbis virus 49 and 26 S molecules contain poly(A); a reproducible fraction of these molecules will not bind to oligo(dT)-cellulose, however, and contains little poly(A). The oligo(dT)-cellulose unbound fraction from virion RNA contains a discrete species of RNA, which does not appear to be due to random degradation, as well as fragments of degraded RNA; this RNA is only 10 to 20% as infectious as oligo(dT)-cellulose-bound or whole virion RNA, suggesting that the poly(A) is essential for replication of the virus. Poly(A) with a size distribution similar to that found in the single-stranded RNAs is also found in Sindbis virus double-stranded RNA species. RF I and RF III contain poly(A) while RF II does not. Stretches of polyuridylic acid (poly(U)) are also found in Sindbis virus double-stranded RNA. This poly(U) is on the minus strand, has a size distribution identical to Sindbis virus poly(A), and occurs once per double-stranded molecule. Poly(U) is found in RF I and RF III but not on RF II. This evidence indicates that the poly(A) in Sindbis virus RNA is synthesized by transcription of a poly(U) template on the viral minus strand.

Additional Information

© 1978 Academic Press, Inc. Accepted December 31, 1977. We thank David L. Gard, Edith M. Lenches, and Agnes V. Raviol for their expert technical assistance in preparing the CEF and BHK cultures and in carrying out parts of some of the experiments described. We thank Jeffrey T. Mayne for providing the A. albopictus cell cultures and for helping us label and prepare mosquito cell-grown virus. We also thank Dr. Ellen G. Strauss for her assistance in preparing this manuscript. This investigation was supported by Grant PCM 75-02469 from the National Science Foundation and by Grants GM-66965 and AI-10793 from the U.S. Public Health Service. One of us (T.K.F.) was supported by a National Science Foundation Graduate Fellowship and by National Institutes of Health Training Grant No. 5-T1-GM86-2.

Additional details

Created:
August 19, 2023
Modified:
October 18, 2023