Title

Tat stimulates cotranscriptional capping of HIV mRNA

GSBS Program

Biochemistry & Molecular Pharmacology

UMMS Affiliation

Graduate School of Biomedical Sciences; Department of Biochemistry and Molecular Pharmacology

Date

11-1-2002

Document Type

Article

Medical Subject Headings

DNA-Binding Proteins; Gene Expression Regulation, Viral; Gene Products, tat; HIV Long Terminal Repeat; HIV-1; Hela Cells; Humans; Methylation; Methyltransferases; Nuclear Proteins; Nucleic Acid Conformation; Nucleotidyltransferases; Promoter Regions (Genetics); RNA Caps; RNA Polymerase II; *RNA Processing, Post-Transcriptional; RNA, Viral; Recombinant Fusion Proteins; *Transcription, Genetic; Viral Proteins; tat Gene Products, Human Immunodeficiency Virus

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

Here we investigated how capping and methylation of HIV pre-mRNAs are coupled to Pol II elongation. Stable binding of the capping enzyme (Mce1) and cap methyltransferase (Hcm1) to template-engaged Pol II depends on CTD phosphorylation, but not on nascent RNA. Both Mce1 and Hcm1 travel with Pol II during elongation. The capping and methylation reactions cannot occur until the nascent pre-mRNA has attained a chain length of 19-22 nucleotides. HIV pre-mRNAs are capped quantitatively when elongation complexes are halted at promoter-proximal positions, but capping is much less efficient during unimpeded Pol II elongation. Cotranscriptional capping of HIV mRNA is strongly stimulated by Tat, and this stimulation requires the C-terminal segment of Tat that mediates its direct binding to Mce1. Our findings implicate capping in an elongation checkpoint critical to HIV gene expression.

Rights and Permissions

Citation: Mol Cell. 2002 Sep;10(3):585-97.

Related Resources

Link to article in PubMed

PubMed ID

12408826