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Journal of Virology, October 1998, p. 7807-7814, Vol. 72, No. 10
Department of Molecular Genetics, Albert
Einstein College of Medicine, Bronx, New York 10461
Received 12 May 1998/Accepted 24 June 1998
In addition to their role in reverse transcription, the R-region
sequences of some retroviruses affect viral transcription. The first 28 nucleotides of the R region within the long terminal repeat (LTR) of
the murine type C retrovirus SL3 were predicted to form a stem-loop
structure. We tested whether this structure affected the
transcriptional activity of the viral LTR. Mutations that altered
either side of the stem and thus disrupted base pairing were generated.
These decreased the level of expression of a reporter gene under the
control of viral LTR sequences about 5-fold in transient expression
assays and 10-fold in cells stably transformed with the LTR-reporter
plasmids. We also generated a compensatory mutant in which both the
ascending and descending sides of the stem were mutated such that the
nucleotide sequence was different but the predicted secondary structure
was maintained. Most of the activity of the wild-type SL3 element was
restored in this mutant. Thus, the stem-loop structure was important
for the maximum activity of the SL3 LTR. Primer extension analysis
indicated that the stem-loop structure affected the levels of
cytoplasmic RNA. Nuclear run-on assays indicated that deletion of the R
region had a small effect on transcriptional initiation and no effect on RNA polymerase processivity. Thus, the main effect of the R-region element was on one or more steps that occurred after the template was
transcribed by RNA polymerase. This finding implied that the main
function of the R-region element involved RNA processing. R-region
sequences of human immunodeficiency virus type 1 or mouse mammary tumor
virus could not replace the SL3 element. R-region sequences from an
avian reticuloendotheliosis virus partially substituted for the SL3
sequences. R-region sequences from Moloney murine leukemia virus or
feline leukemia virus did function in place of the SL3 element. Thus,
the R region element appears to be a general feature of the mammalian
type C genus of retroviruses.
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The Secondary Structure of the R Region of a Murine
Leukemia Virus Is Important for Stimulation of Long Terminal
Repeat-Driven Gene Expression


*
Corresponding author. Mailing address: Department of
Molecular Genetics, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-3715. Fax: (718) 430-8778. E-mail: lenz{at}aecom.yu.edu.
Present address: Hoffmann-La Roche Inc., Nutley, NJ 07110.
Present address: Regeneron Pharmaceuticals, Tarrytown, NY 10591.
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