title: Lysine methylation of HIV-1 Tat regulates transcriptional activity of the viral LTR.
abstract: BACKGROUND
  The rate of transcription of the HIV-1 viral genome is mediated by the interaction of the viral protein Tat with the LTR and other transcriptional machinery. These specific interactions can be affected by the state of post-translational modifications on Tat. Previously, we have shown that Tat can be phosphorylated and acetylated in vivo resulting in an increase in the rate of transcription. In the present study, we investigated whether Tat could be methylated on lysine residues, specifically on lysine 50 and 51, and whether this modification resulted in a decrease of viral transcription from the LTR.
  
  
  RESULTS
  We analyzed the association of Tat with histone methyltransferases of the SUV39-family of SET domain containing proteins in vitro. Tat was found to associate with both SETDB1 and SETDB2, two enzymes which exhibit methyltransferase activity. siRNA against SETDB1 transfected into cell systems with both transient and integrated LTR reporter genes resulted in an increase in transcription of the HIV-LTR in the presence of suboptimal levels of Tat. In vitro methylation assays with Tat peptides containing point mutations at lysines 50 and 51 showed an increased incorporation of methyl groups on lysine 51, however, both residues indicated susceptibility for methylation.
  
  
  CONCLUSIONS
  The association of Tat with histone methyltransferases and the ability for Tat to be methylated suggests an interesting mechanism of transcriptional regulation through the recruitment of chromatin remodeling proteins to the HIV-1 promoter.
authors.affiliation: The George Washington University Medical Center, Department of Biochemistry and Molecular Biology, Washington, DC 20037, USA. bcmrvv@gwumc.edu
authors.forename: Rachel
authors.initials: R
authors.lastname: Van Duyne
authors.forename: Rebecca
authors.initials: R
authors.lastname: Easley
authors.forename: Weilin
authors.initials: W
authors.lastname: Wu
authors.forename: Reem
authors.initials: R
authors.lastname: Berro
authors.forename: Caitlin
authors.initials: C
authors.lastname: Pedati
authors.forename: Zachary
authors.initials: Z
authors.lastname: Klase
authors.forename: Kylene
authors.initials: K
authors.lastname: Kehn-Hall
authors.forename: Elizabeth K
authors.initials: EK
authors.lastname: Flynn
authors.forename: David E
authors.initials: DE
authors.lastname: Symer
authors.forename: Fatah
authors.initials: F
authors.lastname: Kashanchi
mesh_terms: D002460:Cell Line; D017930:Genes, Reporter; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D008239:Lysine / Q000235:genetics / Q000378:metabolism*; D008745:Methylation; D011485:Protein Binding; D011496:Protein Methyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D034741:RNA, Small Interfering / Q000235:genetics; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014775:Virus Activation; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D034741:RNA, Small Interfering; D054322:tat Gene Products, Human Immunodeficiency Virus; D011496:Protein Methyltransferases; D011495:Histone-Lysine N-Methyltransferase; C454062:SETDB1 protein, human; D008239:Lysine