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Journal of Virology, April 2003, p. 4060-4069, Vol. 77, No. 7
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.7.4060-4069.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Possible Role of Dimerization in Human Immunodeficiency Virus Type 1 Genome RNA Packaging

Jun-Ichi Sakuragi,1* Shigeharu Ueda,2 Aikichi Iwamoto,3 and Tatsuo Shioda1

Department of Viral Infections,1 Department of Neurovirology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565,2 Division of Infectious Diseases, Advanced Clinical Research Center, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan3

Received 25 July 2002/ Accepted 20 December 2002

The dimer initiation site/dimer linkage sequence (DIS/DLS) region in the human immunodeficiency virus type 1 (HIV-1) RNA genome is suggested to play important roles in various steps of the virus life cycle. However, due to the presence of a putative DIS/DLS region located within the encapsidation signal region (E/psi), it is difficult to perform a mutational analysis of DIS/DLS without affecting the packaging of RNA into virions. Recently, we demonstrated that duplication of the DIS/DLS region in viral RNA caused the production of partially monomeric RNAs in virions, indicating that the region indeed mediated RNA-RNA interaction. We utilized this system to assess the precise location of DIS/DLS in the 5' region of the HIV-1 genome with minimum effect on RNA packaging. We found that the entire lower stem of the U5/L stem-loop was required for packaging, whereas the region important for dimer formation was only 10 bases long within the lower stem of the U5/L stem-loop. The R/U5 stem-loop was required for RNA packaging but was completely dispensable for dimer formation. The SL1 lower stem was important for both dimerization and packaging, but surprisingly, deletion of the palindromic sequence at the top of the loop only partially affected dimerization. These results clearly indicated that the E/psi of HIV-1 is much larger than the DIS/DLS and that the primary DIS/DLS is completely included in the E/psi. Therefore, it is suggested that RNA dimerization is a part of RNA packaging, which requires multiple steps.

* Corresponding author. Mailing address: Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita City, Osaka 565-0871, Japan. Phone: 81-6-6879-8348. Fax: 81-6-6879-8347. E-mail: sakuragi{at}biken.osaka-u.ac.jp.

Journal of Virology, April 2003, p. 4060-4069, Vol. 77, No. 7
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.7.4060-4069.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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