pX, the HBV-encoded coactivator, interacts with components of the transcription machinery and stimulates transcription in a TAF-independent manner.
The X protein of hepatitis B virus (HBV) coactivates activators bearing potent (mostly acidic) activation domains. Here, we investigated the molecular mechanisms of this coactivation. We show that pX interacts with general transcription factors TFIIB and TFIIH, as well as with the potent activation domain of VP16. TFIIB interacts with both pX and VP16 simultaneously. In addition, the RNA polymerase II enzyme itself binds to pX. By reducing the activity of cellular coactivators, through squelching, we intensify the dependence of the activator on pX-mediated coactivation. Squelching is essentially diminished in the presence of pX, both in vivo and in vitro. The target of pX in this activity is the template-bound activator, and not the squelcher. Furthermore, by following transcription in a TAF-deprived reaction, we demonstrate absolute dependence of the activator on the activity of pX. We propose that pX coactivates transcription by substituting cellular coactivators in activator-preinitiation complex interactions.
Haviv
I
I
Department of Molecular Genetics and Virology, The Weizmann Institute of Science, Rehovot, Israel.
Vaizel
D
D
Shaul
Y
Y
D005656:Fungal Proteins / Q000378:metabolism; D006367:HeLa Cells; D006511:Hepatitis B Antigens / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D012319:RNA Polymerase II / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism*; D035581:Transcription Factor TFIIB; D051758:Transcription Factor TFIIH; D014157:Transcription Factors / Q000378:metabolism*; D019473:Transcription Factors, TFII; D014158:Transcription, Genetic; D054334:Viral Regulatory and Accessory Proteins
D005656:Fungal Proteins; C070633:Gal-VP16; D006511:Hepatitis B Antigens; D015534:Trans-Activators; D035581:Transcription Factor TFIIB; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; D051758:Transcription Factor TFIIH; D012319:RNA Polymerase II