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J Gen Virol 49 (1980), 61-69; DOI 10.1099/0022-1317-49-1-61
© 1980 Society for General Microbiology
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Semliki Forest Virus Replication Complex Capable of Synthesizing 42S and 26S Nascent RNA Chains

Peter J. Gomatos1, Leevi Kääriäinen2, Sirkka Keränen2, Marjut Ranki2 and Dorothea L. Sawicki1,*

1 Division of Animal Virology, The Sloan-Kettering Institute, New York, N.Y. 10021, U.S.A.
2 Department of Virology, University of Helsinki, 00290 Helsinki 29, Finland

A complex synthesizing Semliki Forest virus (SFV)-specific RNAs was purified from infected HeLa cells. During purification, the RNA-synthesizing complex was monitored by the presence of RNA chains synthesized during a 1 min pulse in vivo and the ability to synthesize 42S and 26S RNAs in vitro. Finally, the protein composition of the replication complex was analysed. Thirty to 40% of the pulse-labelled RNAs and 10 to 25% of the polymerase activity present in the postnuclear supernatant were recovered in smooth membranes. At this stage of purification single stranded 42S and 26S RNA were synthesized and released from the replication complex in vitro. After treatment of the smooth membrane fraction with Triton X-100 the replication complex was solubilized. When analysed by sucrose gradient centrifugation, the solubilized replication complex distributed heterogeneously. It had reduced RNA polymerase activity, but was still able to synthesize both 42S and 26S nascent RNA chains which were not released from RIs and RFs. The non-structural protein ns70 was the major virus-specified component associated with the replication complex.

* Present address: Department of Microbiology, Medical College of Ohio at Toledo, Toledo, Ohio 43699, U.S.A.

Received 9 November 1979; accepted 31 January 1980.




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Copyright © 1980 by the Society for General Microbiology.