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Molecular and Cellular Biology, December 2003, p. 8762-8772, Vol. 23, No. 23
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.23.8762-8772.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Human Immunodeficiency Virus Type 1 hnRNP A/B-Dependent Exonic Splicing Silencer ESSV Antagonizes Binding of U2AF65 to Viral Polypyrimidine Tracts

Jeffrey K. Domsic,1,{dagger} Yibin Wang,2 Akila Mayeda,3 Adrian R. Krainer,4 and C. Martin Stoltzfus1,2*

Program in Molecular Biology,1 Department of Microbiology, University of Iowa, Iowa City, Iowa 52242,2 Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Miami, Florida 33136,3 Cold Spring Harbor Laboratory, Cold Spring Harbor, New York 117244

Received 2 June 2003/ Returned for modification 8 July 2003/ Accepted 29 August 2003

Human immunodeficiency virus type 1 (HIV-1) exonic splicing silencers (ESSs) inhibit production of certain spliced viral RNAs by repressing alternative splicing of the viral precursor RNA. Several HIV-1 ESSs interfere with spliceosome assembly by binding cellular hnRNP A/B proteins. Here, we have further characterized the mechanism of splicing repression using a representative HIV-1 hnRNP A/B-dependent ESS, ESSV, which regulates splicing at the vpr 3' splice site. We show that hnRNP A/B proteins bound to ESSV are necessary to inhibit E complex assembly by competing with the binding of U2AF65 to the polypyrimidine tracts of repressed 3' splice sites. We further show evidence suggesting that U1 snRNP binds the 5' splice site despite an almost complete block of splicing by ESSV. Possible splicing-independent functions of U1 snRNP-5' splice site interactions during virus replication are discussed.

* Corresponding author. Mailing address: Department of Microbiology, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-7793. Fax: (319) 335-9006. E-mail: marty-stoltzfus{at}uiowa.edu..

{dagger} Present address: Department of Chemistry and Biochemistry, University of Maryland, Baltimore, MD 21250.

Molecular and Cellular Biology, December 2003, p. 8762-8772, Vol. 23, No. 23
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.23.8762-8772.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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