title: The human RVB complex is required for efficient transcription of type I interferon-stimulated genes.
abstract: Type I interferons (IFNs) stimulate transcription through a latent heterotrimeric transcription factor composed of tyrosine-phosphorylated STAT1 and STAT2 and the DNA binding partner IRF9, with STAT2 contributing a critical transactivation domain. Human RVB1 and RVB2, which are highly conserved AAA(+) ATP binding proteins contained in chromatin-remodeling complexes such as Ino80, SNF2-related CBP activator protein (SRCAP), and Tip60/NuA4, interacted with the transactivation domain of STAT2 in the nuclei of IFN-stimulated cells. RNA interference (RNAi) experiments demonstrated that RVB proteins were required for robust activation of IFN-α-stimulated genes (ISGs). The requirement for RVB proteins was specific to IFN-α/STAT2 signaling; transcription of tumor necrosis factor alpha (TNF-α)- and IFN-γ-driven genes was not affected by RVB1 depletion. Using RNAi-based depletion, we assessed the involvement of catalytic subunits of the RVB-containing Tip60, BRD8, Ino80, SRCAP, and URI complexes. No component other than RVB1/2 was uniquely required for ISG induction, suggesting that RVB1/2 functions as part of an as yet unidentified complex. Chromatin immunoprecipitation assays indicated that RVB1/2 was required for recruitment of RNA polymerase II (Pol II) to ISG promoters but was dispensable for STAT2 recruitment to chromatin. We hypothesize that an RVB1/2 chromatin-remodeling complex is required for efficient Pol II recruitment and initiation at ISG promoters and is recruited through interaction with the STAT2 transactivation domain.
authors.affiliation: Departments of Pathology and Microbiology.
authors.forename: Leonid
authors.initials: L
authors.lastname: Gnatovskiy
authors.forename: Paolo
authors.initials: P
authors.lastname: Mita
authors.forename: David E
authors.initials: DE
authors.lastname: Levy
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins; D057809:HEK293 Cells; D006367:HeLa Cells; D051548:Histone Acetyltransferases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology*; D000074443:Lysine Acetyltransferase 5; D013058:Mass Spectrometry; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D034622:RNA Interference; D012319:RNA Polymerase II / Q000378:metabolism; D011988:Receptors, Thyroid Hormone / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050795:STAT2 Transcription Factor / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors; D014158:Transcription, Genetic / Q000187:drug effects*; D000092468:Bromodomain Containing Proteins
chemical_list: D000074183:ATPases Associated with Diverse Cellular Activities; D000251:Adenosine Triphosphatases; D002352:Carrier Proteins; D004265:DNA Helicases; D004268:DNA-Binding Proteins; D051548:Histone Acetyltransferases; D007370:Interferon Type I; D000074443:Lysine Acetyltransferase 5; D012319:RNA Polymerase II; D011988:Receptors, Thyroid Hormone; D050795:STAT2 Transcription Factor; D014157:Transcription Factors; C099139:BRD8 protein, human; D000092468:Bromodomain Containing Proteins; C098489:KAT5 protein, human; C524421:INO80 protein, human; C119652:SRCAP protein, human; C486549:RUVBL1 protein, human; C122079:RUVBL2 protein, human