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RNA splicing at human immunodeficiency virus type 1 3' splice site A2 is regulated by binding of hnRNP A/B proteins to an exonic splicing silencer element.
J Virol. 2001 Sep; 75(18):8487-97.JV

Abstract

The synthesis of human immunodeficiency virus type 1 (HIV-1) mRNAs is a complex process by which more than 30 different mRNA species are produced by alternative splicing of a single primary RNA transcript. HIV-1 splice sites are used with significantly different efficiencies, resulting in different levels of mRNA species in infected cells. Splicing of Tat mRNA, which is present at relatively low levels in infected cells, is repressed by the presence of exonic splicing silencers (ESS) within the two tat coding exons (ESS2 and ESS3). These ESS elements contain the consensus sequence PyUAG. Here we show that the efficiency of splicing at 3' splice site A2, which is used to generate Vpr mRNA, is also regulated by the presence of an ESS (ESSV), which has sequence homology to ESS2 and ESS3. Mutagenesis of the three PyUAG motifs within ESSV increases splicing at splice site A2, resulting in increased Vpr mRNA levels and reduced skipping of the noncoding exon flanked by A2 and D3. The increase in Vpr mRNA levels and the reduced skipping also occur when splice site D3 is mutated toward the consensus sequence. By in vitro splicing assays, we show that ESSV represses splicing when placed downstream of a heterologous splice site. A1, A1(B), A2, and B1 hnRNPs preferentially bind to ESSV RNA compared to ESSV mutant RNA. Each of these proteins, when added back to HeLa cell nuclear extracts depleted of ESSV-binding factors, is able to restore splicing repression. The results suggest that coordinate repression of HIV-1 RNA splicing is mediated by members of the hnRNP A/B protein family.

Authors+Show Affiliations

Department of Microbiology, University of Iowa, Iowa City, Iowa 52242, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11507194

Citation

Bilodeau, P S., et al. "RNA Splicing at Human Immunodeficiency Virus Type 1 3' Splice Site A2 Is Regulated By Binding of hnRNP A/B Proteins to an Exonic Splicing Silencer Element." Journal of Virology, vol. 75, no. 18, 2001, pp. 8487-97.
Bilodeau PS, Domsic JK, Mayeda A, et al. RNA splicing at human immunodeficiency virus type 1 3' splice site A2 is regulated by binding of hnRNP A/B proteins to an exonic splicing silencer element. J Virol. 2001;75(18):8487-97.
Bilodeau, P. S., Domsic, J. K., Mayeda, A., Krainer, A. R., & Stoltzfus, C. M. (2001). RNA splicing at human immunodeficiency virus type 1 3' splice site A2 is regulated by binding of hnRNP A/B proteins to an exonic splicing silencer element. Journal of Virology, 75(18), 8487-97.
Bilodeau PS, et al. RNA Splicing at Human Immunodeficiency Virus Type 1 3' Splice Site A2 Is Regulated By Binding of hnRNP A/B Proteins to an Exonic Splicing Silencer Element. J Virol. 2001;75(18):8487-97. PubMed PMID: 11507194.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - RNA splicing at human immunodeficiency virus type 1 3' splice site A2 is regulated by binding of hnRNP A/B proteins to an exonic splicing silencer element. AU - Bilodeau,P S, AU - Domsic,J K, AU - Mayeda,A, AU - Krainer,A R, AU - Stoltzfus,C M, PY - 2001/8/17/pubmed PY - 2001/9/28/medline PY - 2001/8/17/entrez SP - 8487 EP - 97 JF - Journal of virology JO - J Virol VL - 75 IS - 18 N2 - The synthesis of human immunodeficiency virus type 1 (HIV-1) mRNAs is a complex process by which more than 30 different mRNA species are produced by alternative splicing of a single primary RNA transcript. HIV-1 splice sites are used with significantly different efficiencies, resulting in different levels of mRNA species in infected cells. Splicing of Tat mRNA, which is present at relatively low levels in infected cells, is repressed by the presence of exonic splicing silencers (ESS) within the two tat coding exons (ESS2 and ESS3). These ESS elements contain the consensus sequence PyUAG. Here we show that the efficiency of splicing at 3' splice site A2, which is used to generate Vpr mRNA, is also regulated by the presence of an ESS (ESSV), which has sequence homology to ESS2 and ESS3. Mutagenesis of the three PyUAG motifs within ESSV increases splicing at splice site A2, resulting in increased Vpr mRNA levels and reduced skipping of the noncoding exon flanked by A2 and D3. The increase in Vpr mRNA levels and the reduced skipping also occur when splice site D3 is mutated toward the consensus sequence. By in vitro splicing assays, we show that ESSV represses splicing when placed downstream of a heterologous splice site. A1, A1(B), A2, and B1 hnRNPs preferentially bind to ESSV RNA compared to ESSV mutant RNA. Each of these proteins, when added back to HeLa cell nuclear extracts depleted of ESSV-binding factors, is able to restore splicing repression. The results suggest that coordinate repression of HIV-1 RNA splicing is mediated by members of the hnRNP A/B protein family. SN - 0022-538X UR - https://www.unboundmedicine.com/medline/citation/11507194/RNA_splicing_at_human_immunodeficiency_virus_type_1_3'_splice_site_A2_is_regulated_by_binding_of_hnRNP_A/B_proteins_to_an_exonic_splicing_silencer_element_ L2 - http://jvi.asm.org/cgi/pmidlookup?view=long&pmid=11507194 DB - PRIME DP - Unbound Medicine ER -