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Journal of Virology, August 2002, p. 8019-8030, Vol. 76, No. 16
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.16.8019-8030.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

RelA-Associated Inhibitor Blocks Transcription of Human Immunodeficiency Virus Type 1 by Inhibiting NF-{kappa}B and Sp1 Actions

Norio Takada,1,2 Takaomi Sanda,1 Hiroshi Okamoto,1 Jian-Ping Yang,1 Kaori Asamitsu,1 Lilen Sarol,1 Genjiro Kimura,2 Hiroaki Uranishi,1 Toshifumi Tetsuka,1 and Takashi Okamoto1*

Department of Molecular Genetics,1 3rd Department of Internal Medicine, Nagoya City University Medical School, Nagoya 467-8601, Japan2

Received 28 January 2002/ Accepted 23 April 2002

RelA-associated inhibitor (RAI) is an inhibitor of nuclear factor {kappa}B (NF-{kappa}B) newly identified by yeast two-hybrid screen as an interacting protein of the p65 (RelA) subunit. In this study, we attempted to examine the effect of RAI on transcription and replication of human immunodeficiency virus type 1 (HIV-1). We found that RAI inhibited gene expression from the HIV-1 long terminal repeat (LTR) even at the basal level. Upon in vitro DNA-binding reactions, RAI could directly block the DNA-binding of p65 subunit of NF-{kappa}B but not that of the p50 subunit or AP1. We found that RAI could also inhibit the DNA-binding of Sp1 and thus inhibit the basal HIV-1 promoter activity. We further examined the effects of RAI on Sp1 and found that RAI colocalizes with Sp1 in the nucleus and interacts with Sp1 in vitro and in vivo. Moreover, we found that RAI efficiently blocked the HIV-1 replication when cotransfected with a full-length HIV-1 clone. These findings indicate that RAI acts as an efficient inhibitor of HIV-1 gene expression in which both NF-{kappa}B and Sp1 play major roles.

* Corresponding author. Mailing address: Department of Molecular Genetics, Nagoya City University Medical School, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, Aichi 467-8601, Japan. Phone: 81-52-853-8204. Fax: 81-52-859-1235. E-mail: tokamoto{at}med.nagoya-cu.ac.jp.

Journal of Virology, August 2002, p. 8019-8030, Vol. 76, No. 16
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.16.8019-8030.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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