Interaction of protein inhibitor of activated STAT (PIAS) proteins with the TATA-binding protein, TBP.
Transcription activators often recruit promoter-targeted assembly of a pre-initiation complex; many repressors antagonize recruitment. These activities can involve direct interactions with proteins in the pre-initiation complex. We used an optimized yeast two-hybrid system to screen mouse pregnancy-associated libraries for proteins that interact with TATA-binding protein (TBP). Screens revealed an interaction between TBP and a single member of the zinc finger family of transcription factors, ZFP523. Two members of the protein inhibitor of activated STAT (PIAS) family, PIAS1 and PIAS3, also interacted with TBP in screens. Endogenous PIAS1 and TBP co-immunoprecipitated from nuclear extracts, suggesting the interaction occurred in vivo. In vitro-translated PIAS1 and TBP co-immunoprecipitated, which indicated that other nuclear proteins were not required for the interaction. Deletion analysis mapped the PIAS-interacting domain of TBP to the conserved TBP(CORE) and the TBP-interacting domain on PIAS1 to a 39-amino acid C-terminal region. Mammals issue seven known PIAS proteins from four pias genes, pias1, pias3, piasx, and piasy, each with different cell type-specific expression patterns; the TBP-interacting domain reported here is the only part of the PIAS C-terminal region shared by all seven PIAS proteins. Direct analyses indicated that PIASx and PIASy also interacted with TBP. Our results suggest that all PIAS proteins might mediate situation-specific regulatory signaling at the TBP interface and that previously unknown levels of complexity could exist in the gene regulatory interplay between TBP, PIAS proteins, ZFP523, and other transcription factors.
Prigge
Justin R
JR
Veterinary Molecular Biology, Montana State University, Bozeman, Montana 59717, USA.
Schmidt
Edward E
EE
D000595:Amino Acid Sequence; D000818:Animals; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry; D050828:Protein Inhibitors of Activated STAT / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D035181:TATA-Box Binding Protein / Q000378:metabolism / Q000502:physiology*; D014157:Transcription Factors / Q000737:chemistry*; D015533:Transcriptional Activation; D044767:Ubiquitin-Protein Ligases
D009687:Nuclear Proteins; C494211:Pias1 protein, mouse; C109279:Pias3 protein, mouse; C414349:Piasy protein, mouse; D050828:Protein Inhibitors of Activated STAT; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; C511247:Zfp523 protein, mouse; C485521:Miz1 protein, mouse; D044767:Ubiquitin-Protein Ligases