Interaction of human immunodeficiency virus type 1 Tat with a unique site of TFIID inhibits negative cofactor Dr1 and stabilizes the TFIID-TFIIA complex.
We have previously reported the direct physical interaction between the human immunodeficiency virus (HIV) type I Tat protein and the basal transcription factor TBP/TFIID. Affinity chromatography demonstrated that wild-type Tat, but not a transactivation mutant of Tat, was capable of depleting TBP/TFIID from cell extracts. These experiments represented the first demonstration of a basal transcription factor that binds, in an activation-dependent manner, to Tat. We now report that the Tat-TBP interaction can be detected in HIV type 1-infected cells. The domain of TBP interacting with Tat has been mapped from amino acids 163 to 196 by using deletion and site-specific mutants of TBP. This domain of TBP, which includes the HI and S2 domains, is distinct from the H2 binding site for other activator proteins, such as E1A. The interaction of Tat with TFIID regulates the binding of accessory proteins to TFIID. Tat stabilizes the interaction of TFIID with TFIIA in a gel shift assay. In addition, Tat competes for Dr1 interaction with TBP. Our results suggest that the basal transcription factor TBP/TFIID represents an important regulatory molecule in HIV transcription.
Kashanchi
F
F
Laboratory of Molecular Virology, National Cancer Institute, Bethesda, Maryland 20892, USA.
Khleif
S N
SN
Duvall
J F
JF
Sadaie
M R
MR
Radonovich
M F
MF
Cho
M
M
Martin
M A
MA
Chen
S Y
SY
Weinmann
R
R
Brady
J N
JN
D002460:Cell Line; D015696:Gene Products, tat / Q000378:metabolism*; D015658:HIV Infections / Q000378:metabolism / Q000821:virology*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D035561:Transcription Factor TFIIA; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000378:metabolism*; D054322:tat Gene Products, Human Immunodeficiency Virus
D015696:Gene Products, tat; D010750:Phosphoproteins; D035561:Transcription Factor TFIIA; D035362:Transcription Factor TFIID; D014157:Transcription Factors; C075995:down-regulator of transcription 1; D054322:tat Gene Products, Human Immunodeficiency Virus