The HIV-1 Tat Protein Enhances Splicing at the Major Splice Donor Site

• Published in: Journal of virology Vol. 92; no. 14
• Main Authors: Mueller, Nancy, Pasternak, Alexander O, Klaver, Bep, Cornelissen, Marion, Berkhout, Ben, Das, Atze T
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 15.07.2018
• Subjects: Gene Expression Regulation, Viral; Gene Products, tat - genetics; Gene Products, tat - metabolism; Genome and Regulation of Viral Gene Expression; HEK293 Cells; HIV Infections - virology; HIV-1 - genetics; HIV-1 - isolation & purification; Humans; RNA Splicing; RNA, Viral - genetics; Virus Replication; human immunodeficiency virus; Tat protein; transcription; VP16; RNA splicing
• ISSN: 0022-538X; 1098-5514
• DOI: 10.1128/jvi.01855-17

Transcription of the HIV-1 proviral DNA and subsequent processing of the primary transcript results in the production of a large set of unspliced and differentially spliced viral RNAs. The major splice donor site (5'ss) that is located in the untranslated leader of the HIV-1 transcript is used for the production of all spliced RNAs, and splicing at this site has to be tightly regulated to allow the balanced production of all viral RNAs and proteins. We demonstrate that the viral Tat protein, which is known to activate viral transcription, also stimulates splicing at the major 5'ss. As for the transcription effect, Tat requires the viral long terminal repeat promoter and the -acting responsive RNA hairpin for splicing regulation. These results indicate that HIV-1 transcription and splicing are tightly coupled processes through the coordinated action of the essential Tat protein. The HIV-1 proviral DNA encodes a single RNA transcript that is used as RNA genome and packaged into newly assembled virus particles. This full-length RNA is also used as mRNA for the production of structural and enzymatic proteins. Production of other essential viral proteins depends on alternative splicing of the primary transcript, which yields a large set of differentially spliced mRNAs. Optimal virus replication requires a balanced production of all viral RNAs, which means that the splicing process has to be strictly regulated. We show that the HIV-1 Tat protein, a factor that is well known for its transcription activating function, also stimulates splicing. Thus, Tat controls not only the level of the viral RNA but also the balance between spliced and unspliced RNAs.