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Journal of Virology, March 2001, p. 2557-2565, Vol. 75, No. 6
McArdle Laboratory for Cancer Research,
University of Wisconsin-Madison, Madison, Wisconsin
537061; Department of Viral Infections,
Research Institute for Microbial Diseases, Osaka University, Suita,
Osaka 565-0871, Japan2; and
Department of Molecular Genetics and Microbiology,
University of New Mexico Health Science Center, Albuquerque, New
Mexico 87131-50013
Received 11 September 2000/Accepted 19 December 2000
The dimerization initiation site (DIS) and the dimer linkage
sequences (DLS) of human immunodeficiency virus type 1 have been shown
to mediate in vitro dimerization of genomic RNA. However, the precise
role of the DIS-DLS region in virion assembly and RNA dimerization in
virus particles has not been fully elucidated, since deletion or
mutation of the DIS-DLS region also abolishes the packaging ability of
genomic RNA. To characterize the DIS-DLS region without altering
packaging ability, we generated mutant constructs carrying a
duplication of approximately 1,000 bases including the encapsidation
signal and DIS-DLS (E/DLS) region. We found that duplication of the
E/DLS region resulted in the appearance of monomeric RNA in virus
particles. No monomers were observed in virions of mutants carrying the
E/DLS region only at ectopic positions. Monomers were not observed when
pol or env regions were duplicated, indicating
an absolute need for two intact E/DLS regions on the same RNA for
generating particles with monomeric RNA. These monomeric RNAs were most
likely generated by intramolecular interaction between two E/DLS
regions on one genome. Moreover, incomplete genome dimerization did not
affect RNA packaging and virion formation. Examination of
intramolecular interaction between E/DLS regions could be a convenient
tool for characterizing the E/DLS region in virion assembly and RNA
dimerization within virus particles.
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.6.2557-2565.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Duplication of the Primary Encapsidation and Dimer
Linkage Region of Human Immunodeficiency Virus Type 1 RNA Results in
the Appearance of Monomeric RNA in Virions
*
Corresponding author. Mailing address: Department of
Viral Infections, Research Institute for Microbial Diseases, Osaka
University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan. Phone:
81-6-6879-8348. Fax: 81-6-6879-8347. E-mail:
sakuragi{at}biken.osaka-u.ac.jp.
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