The HIV-1 Vpr co-activator induces a conformational change in TFIIB.
Vpr is a HIV-1 virion-associated protein which plays a role in viral replication and in transcription and cell proliferation. We have previously reported that Vpr stimulates transcription of genes lacking a common DNA target sequence likely through its ability to interact with TFIIB. However, the molecular mechanism of the Vpr-mediated transcription remains to be precisely defined. In this in vitro study, we show that the binding site of Vpr in TFIIB overlaps the domain of TFIIB which is engaged in the intramolecular bridge between the N- and C-terminus of TFIIB, highly suggesting that binding of Vpr may induce a change in the conformation of TFIIB. Indeed, with a partial proteolysis assay using V8 protease, we demonstrate that Vpr has the ability to change the conformation of TFIIB. We investigated in this partial proteolysis assay a series of Vpr-mutated proteins previously defined for their transactivation properties. Our data show a correlation between the ability of Vpr-mutated proteins to stimulate transcription and their ability to induce a conformational change in TFIIB, indicating a functional relevance of the Vpr-TFIIB interaction.
Agostini
I
I
INSERM U372, Marseille, France.
Navarro
J M
JM
Bouhamdan
M
M
Willetts
K
K
Rey
F
F
Spire
B
B
Vigne
R
R
Pomerantz
R
R
Sire
J
J
D001665:Binding Sites; D016336:Gene Products, vpr / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D035581:Transcription Factor TFIIB; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D054325:vpr Gene Products, Human Immunodeficiency Virus
D016336:Gene Products, vpr; D011993:Recombinant Fusion Proteins; D035581:Transcription Factor TFIIB; D014157:Transcription Factors; D054325:vpr Gene Products, Human Immunodeficiency Virus