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J Virol, April 1998, p. 3407-3411, Vol. 72, No. 4
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Mutational Analysis of the Rous Sarcoma Virus DR Posttranscriptional Control Element

Robert A. Ogertdagger and Karen L. Beemon*

Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218

Received 28 July 1997/Accepted 3 December 1997

The direct repeat (DR) sequences flanking the src gene in Rous sarcoma virus are essential posttranscriptional control elements; at least one copy of this sequence is necessary for cytoplasmic accumulation of unspliced viral RNA. These sequences promote Rev-independent human immunodeficiency virus type 1 expression, suggesting they act as constitutive transport elements (CTEs). To determine which regions of this sequence are critical for CTE function, mutations in the downstream DR were generated and tested in a viral deletion construct lacking src and the upstream DR. Two single-point mutations and three different clustered mutations caused substantial reductions in reverse transcriptase activity, Gag protein levels, and unspliced viral RNA in the cytoplasm. Three conserved regions of the CTE, including nucleotides 8844 to 8847, 8862 to 8864, and 8868 to 8870, were most sensitive to inactivation by mutagenesis.

* Corresponding author. Mailing address: Department of Biology, Johns Hopkins University, 3400 N. Charles St., Baltimore, MD 21218. Phone: (410) 516-7292. Fax: (410) 516-7289. E-mail: KLB{at}jhu.edu.

dagger Present address: Laboratory of Molecular Microbiology, NIAID, NIH, Bethesda, MD 20892.




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