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Journal of Virology, June 2000, p. 5729-5735, Vol. 74, No. 12
McGill AIDS Centre, Lady Davis Institute for
Medical Research, Sir Mortimer B. Davis-Jewish General
Hospital,1 and Department of
Medicine2 and Department of Microbiology
and Immunology,3 McGill University, Montreal,
Quebec, Canada H3T 1E2
Received 23 September 1999/Accepted 17 March 2000
The kissing-loop domain (KLD) encompasses a stem-loop, named
kissing-loop or dimerization initiation site (DIS) hairpin (nucleotides [nt] 248 to 270 in the human immunodeficiency virus type 1 strains HIV-1Lai and HIV-1Hxb2), seated on top of a
12-nt stem-internal loop called stem-loop B (nt 243 to 247 and 271 to
277). Destroying stem-loop B reduced genome dimerization by ~50% and
proviral DNA synthesis by ~85% and left unchanged the dissociation
temperature of dimeric genomic RNA. The most affected step of reverse
transcription was plus-strand DNA transfer, which was reduced by
~80%. Deleting nt 241 to 256 or 200 to 256 did not reduce genome
dimerization significantly more than the destruction of stem-loop B or
the DIS hairpin. We conclude that the KLD is nonmodular: mutations in
stem-loop B and in the DIS hairpin have similar effects on genome
dimerization, reverse transcription, and encapsidation and are also
"nonadditive"; i.e., a larger deletion spanning both of these
structures has the same effects on genome dimerization and
encapsidation as if stem-loop B strongly impacted DIS hairpin function
and vice versa. A C258G transversion in the palindrome of the
kissing-loop reduced genome dimerization by ~50% and viral infectivity by ~1.4 log. Two mutations, CGCG261
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Impact of Human Immunodeficiency Virus Type 1 RNA Dimerization on
Viral Infectivity and of Stem-Loop B on RNA Dimerization and
Reverse Transcription and Dissociation of Dimerization from
Packaging
UUAA261
(creating a weaker palindrome) and a
241-256 suppressor mutation,
were each able to reduce genome dimerization but leave genome packaging unaffected.
*
Corresponding author. Mailing address: Lady Davis
Institute for Medical Research, 3755 Cote Ste. Catherine Rd., Montreal, Quebec, Canada H3T 1E2. Phone: (514) 340-8260. Fax: (514) 340-7502. E-mail: laughrea{at}hotmail.com.
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