Molecular basis of CD4 repression by the Swi/Snf-like BAF chromatin remodeling complex.
The Brg1/Brm-associated factor (BAF) chromatin remodeling complex directly binds the CD4 silencer and is essential for CD4 repression during T-cell development, because deletion of the ATPase subunit Brg1 or a dominant negative mutant of BAF57 each impairs CD4 repression in early thymocytes. Paradoxically, BAF57 is dispensable for remodeling nucleosomes in vitro or for binding of the BAF complex to the CD4 silencer in vivo. Thus, it is unclear whether BAF57-dependent CD4 repression involves chromatin remodeling and, if so, how the remodeling translates into CD4 repression. Here we show that nucleosomes at the CD4 silencer occupy multiple translational frames. BAF57 dominant negative mutant does not alter these frames, but reduces the accessibility of the entire silencer without affecting the flanking regions, concomitant with localized accumulation of linker histone H1 and eviction of Runx1, a key repressor of CD4 transcription that directly binds the CD4 silencer. Our data indicate that precise nucleosome positioning is not critical for the CD4 silencer function and that BAF57 participates in remodeling H1-containing chromatin at the CD4 silencer, which enables Runx1 to access the silencer and repress CD4. In addition to BAF57, multiple other subunits in the BAF complex are also dispensable for chromatin remodelling in vitro. Our data suggest that these subunits could also help remodel chromatin at a step after the recruitment of the BAF complex to target genes.
Wan
Mimi
M
Department of Immunobiology, Yale University Medical School, New Haven, CT 06520, USA.
Zhang
Jianmin
J
Lai
Dazhi
D
Jani
Anant
A
Prestone-Hurlburt
Paula
P
Zhao
Lulu
L
Ramachandran
Aruna
A
Schnitzler
Gavin R
GR
Chi
Tian
T
D000818:Animals; D015704:CD4 Antigens / Q000235:genetics*; D002843:Chromatin / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D050676:Core Binding Factor Alpha 2 Subunit / Q000378:metabolism; D005786:Gene Expression Regulation; D006657:Histones / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation / Q000235:genetics; D037941:Silencer Elements, Transcriptional; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
D015704:CD4 Antigens; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D050676:Core Binding Factor Alpha 2 Subunit; D006657:Histones; C493726:Runx1 protein, mouse; C445179:SWI-SNF-B chromatin-remodeling complex; C465207:Smarce1 protein, mouse; D014157:Transcription Factors