PRMT6 diminishes HIV-1 Rev binding to and export of viral RNA

Retrovirology. 2006 Dec 18;3:93. doi: 10.1186/1742-4690-3-93.

Abstract

Background: The HIV-1 Rev protein mediates nuclear export of unspliced and partially spliced viral RNA through interaction with the Rev response element (RRE) by means of an arginine rich motif that is similar to the one found in Tat. Since Tat is known to be asymmetrically arginine dimethylated by protein arginine methyltransferase 6 (PRMT6) in its arginine rich motif, we investigated whether the Rev protein could act as a substrate for this enzyme.

Results: Here, we report the methylation of Rev due to a single arginine dimethylation in the N-terminal portion of its arginine rich motif and the association of Rev with PRMT6 in vivo. Further analysis demonstrated that the presence of increasing amounts of wild-type PRMT6, as well as a methylation-inactive mutant PRMT6, dramatically down-regulated Rev protein levels in concentration-dependent fashion, which was not dependent on the methyltransferase activity of PRMT6. Quantification of Rev mRNA revealed that attenuation of Rev protein levels was due to a posttranslational event, carried out by a not yet defined activity of PRMT6. However, no relevant protein attenuation was observed in subsequent chloramphenicol acetyltransferase (CAT) expression experiments that screened for RNA export and interaction with the RRE. Binding of the Rev arginine rich motif to the RRE was reduced in the presence of wild-type PRMT6, whereas mutant PRMT6 did not exert this negative effect. In addition, diminished interactions between viral RNA and mutant Rev proteins were observed, due to the introduction of single arginine to lysine substitutions in the Rev arginine rich motif. More importantly, wild-type PRMT6, but not mutant methyltransferase, significantly decreased Rev-mediated viral RNA export from the nucleus to the cytoplasm in a dose-dependent manner.

Conclusion: These findings indicate that PRMT6 severely impairs the function of HIV-1 Rev.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus
  • Cell Nucleus / metabolism*
  • Gene Expression Regulation, Viral*
  • Gene Products, rev / genetics
  • Gene Products, rev / metabolism*
  • Genes, rev
  • HIV-1 / genetics
  • HIV-1 / metabolism*
  • HeLa Cells
  • Humans
  • Methylation
  • Nuclear Proteins / metabolism*
  • Protein-Arginine N-Methyltransferases / metabolism*
  • RNA, Viral / genetics
  • RNA, Viral / metabolism*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism
  • rev Gene Products, Human Immunodeficiency Virus

Substances

  • Gene Products, rev
  • Nuclear Proteins
  • RNA, Viral
  • RNA-Binding Proteins
  • rev Gene Products, Human Immunodeficiency Virus
  • PRMT6 protein, human
  • Protein-Arginine N-Methyltransferases