ArrayExpress Uploader0mouse8818arrayexpress_sid8The composition of chromatin remodeling complexes dictates how these enzymes control transcriptional programs and cellular identity....18809673/profile/8773/arrayexpressuploader04chromatinproteinDec.12, 2014Falsebrd7-a-novel-pbaf-specific-swisnf-subunit-is-requihttp://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-7791E-GEOD-7791_A-AFFY-36Brd7, a novel PBAF-specific SWI/SNF subunit, is required for gene activation and repression in embryonic stem cellsNov.24, 2014The composition of chromatin remodeling complexes dictates how these enzymes control transcriptional programs and cellular identity. Here, we investigate the composition of SWI/SNF complexes in embryonic stem cells (ESCs). In contrast to differentiated cells, ESCs have a biased incorporation of certain paralogous SWI/SNF subunits, with low levels of Brm, BAF170 and ARID1B. Upon differentiation, the expression of these subunits increases, resulting in a higher diversity of compositionally distinct SWI/SNF enzymes. We also identify Brd7 as a novel component of the PBAF complex in both ESCs and differentiated cells. Using shRNA-mediated depletion of Brg1, we show that SWI/SNF can function as both a repressor and an activator in pluripotent cells, regulating expression of developmental modifiers and signaling components such as Nodal, ADAMTS1, Bmi-1, CRABP1 and TRH. Knock-down studies of PBAF-specific Brd7 and of a signature subunit within the BAF complex, ARID1A, show that these two sub-complexes affect SWI/SNF target genes differentially, in some cases even antagonistically. This may be due to their different biochemical properties. Finally, we examine the role of SWI/SNF in regulating its target genes during differentiation. We find that SWI/SNF affects recruitment of components of the pre-initiation complex in a promoter-specific manner, to modulate transcription positively or negatively. Taken together, our results provide insight into the function of compositionally diverse SWI/SNF enzymes that underlie their inherent gene-specific mode of action. R1 ESCs were infected in duplicates with shRNA targeting Brg1 or GLUT4 (as a control). Knockdown of Brg1 mRNA affected Brg1 protein levels efficiently. RNA was isolated 67 hours post-infection and analyzed using microarrays.E-GEOD-7791http://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-7791/samples/