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J. Biol. Chem., Vol. 276, Issue 39, 36482-36492, September 28, 2001
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From the The role of RNA-RNA template interactions in
facilitating recombination during reverse transcription of minus strand
DNA has been examined. The tested hypothesis is that template switching by reverse transcriptase is promoted at sites where homologous regions
of two RNAs are brought in close proximity via stable intertemplate
interactions. Frequency and distribution of template switching between
homologous donor and acceptor RNAs were examined within the human
immunodeficiency virus type I (HIV-I) 5'-untranslated region (UTR)
containing the dimer initiation sequence (DIS). Results were compared
with control nondimerizing templates from the pol region. The
dimerizing UTR templates displayed a 4-fold higher transfer efficiency
than the control. A striking 53% of transfers in the UTR mapped near
the DIS, of which two-thirds occurred immediately 5' to this sequence.
In the UTR template, deletion of the DIS hairpin disrupted template
dimerization and caused a 4-fold drop in transfer efficiency. Insertion
of the DIS within the pol template increased both dimerization and
transfer efficiency. Transfer distributions revealed that in both sets
of templates, DIS-induced dimerization increased the efficiency of
transfers across the whole template, with the transfers peaking around
the dimerization site. Overall, these results suggest that
template dimerization facilitated by the unique geometry of the
DIS-promoted kissing interactions effectively promotes recombination
within the HIV-I 5'-UTR.
The Kissing Hairpin Sequence Promotes Recombination within the
HIV-I 5' Leader Region*
,, and§¶
Department of Biochemistry and Biophysics
and § Cancer Center, University of Rochester Medical Center,
Rochester, New York 14642
*
This work was supported by National Institutes of Health
Grant GM 49573 (to R. A. B. and P. J. F.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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