Extensive interactions between HIV TAT and TAF(II)250.
The HIV transactivator, Tat, has been shown to be capable of potent repression of transcription initiation. Repression is mediated by the C-terminal segment of Tat, which binds the TFIID component, TAF(II)250, although the site(s) of interaction were not defined previously. We now report that the interaction between Tat and TAF(II)250 is extensive and involves multiple contacts between the Tat protein and TAF(II)250. The C-terminal domain of Tat, which is necessary for repression of transcription initiation, binds to a segment of TAF(II)250 that encompasses its acetyl transferase (AT) domain (885-1034 amino acids (aa)). Surprisingly, the N-terminal segment of Tat, which contains its activation domains, also binds to TAF(II)250 and interacts with two discontinuous segments of TAF(II)250 located between 885 and 984 aa and 1120 and 1279 aa. Binding of Tat to the 885-984 aa segment of TAF(II)250 requires the cysteine-rich domain of Tat, but not the acidic or glutamine-rich domains. Binding by the N-terminal domain of Tat to the 1120-1279 aa TAF(II)250 segment does not involve the acidic, cysteine- or glutamine-rich domains. Repression of transcription initiation by Tat requires functional TAF(II)250. We now demonstrate that transcription of the HIV LTR does not depend on TAF(II)250 which may account for its resistance to Tat mediated repression.
Experimental Immunology Branch, NCI, NIH, 9000 Rockville Pike, Bethesda, MD 20892, USA. jw35s@nih.gov
J D
JD
Weissman
J R
JR
Hwang
D S
DS
Singer
D000123:Acetyltransferases / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D003545:Cysteine / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000737:chemistry*; D015696:Gene Products, tat / Q000737:chemistry*; D006678:HIV / Q000235:genetics*; D016325:HIV Long Terminal Repeat / Q000235:genetics; D051548:Histone Acetyltransferases; D008958:Models, Molecular; D009687:Nuclear Proteins / Q000737:chemistry*; D011401:Promoter Regions, Genetic; D035381:TATA-Binding Protein Associated Factors; D015534:Trans-Activators / Q000737:chemistry*; D035362:Transcription Factor TFIID; D014158:Transcription, Genetic; D014162:Transfection; D054322:tat Gene Products, Human Immunodeficiency Virus
D004268:DNA-Binding Proteins; D015696:Gene Products, tat; D009687:Nuclear Proteins; D035381:TATA-Binding Protein Associated Factors; D015534:Trans-Activators; D035362:Transcription Factor TFIID; D054322:tat Gene Products, Human Immunodeficiency Virus; D000123:Acetyltransferases; D051548:Histone Acetyltransferases; C069001:TATA-binding protein associated factor 250 kDa; D003545:Cysteine