title: SAF-A forms a complex with BRG1 and both components are required for RNA polymerase II mediated transcription.
abstract: BACKGROUND
  Scaffold attachment factor A (SAF-A) participates in the regulation of gene expression by organizing chromatin into transcriptionally active domains and by interacting directly with RNA polymerase II.
  
  
  METHODS
  Here we use co-localization, co-immunoprecipitation (co-IP) and in situ proximity ligation assay (PLA) to identify Brahma Related Gene 1 (BRG1), the ATP-driven motor of the human SWI-SNF chromatin remodeling complex, as another SAF-A interaction partner in mouse embryonic stem (mES) cells. We also employ RNA interference to investigate functional aspects of the SAF-A/BRG1 interaction.
  
  
  RESULTS
  We find that endogenous SAF-A protein interacts with endogenous BRG1 protein in mES cells, and that the interaction does not solely depend on the presence of mRNA. Moreover the interaction remains intact when cells are induced to differentiate. Functional analyses reveal that dual depletion of SAF-A and BRG1 abolishes global transcription by RNA polymerase II, while the nucleolar RNA polymerase I transcription machinery remains unaffected.
  
  
  CONCLUSIONS
  We demonstrate that SAF-A interacts with BRG1 and that both components are required for RNA Polymerase II Mediated Transcription.
authors.affiliation: Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Dzeneta
authors.initials: D
authors.lastname: Vizlin-Hodzic
authors.forename: Rikard
authors.initials: R
authors.lastname: Runnberg
authors.forename: Jessica
authors.initials: J
authors.lastname: Ryme
authors.forename: Stina
authors.initials: S
authors.lastname: Simonsson
authors.forename: Tomas
authors.initials: T
authors.lastname: Simonsson
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004265:DNA Helicases / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology; D034722:Heterogeneous-Nuclear Ribonucleoprotein U / Q000378:metabolism*; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D008954:Models, Biological; D009687:Nuclear Proteins / Q000378:metabolism*; D011485:Protein Binding; D012319:RNA Polymerase II / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D034722:Heterogeneous-Nuclear Ribonucleoprotein U; D009687:Nuclear Proteins; D012333:RNA, Messenger; C499336:SMARCA1 protein, human; D014157:Transcription Factors; D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases; C084108:SMARCA4 protein, human; D004265:DNA Helicases