Inhibition of chromatin remodeling by polycomb group protein posterior sex combs is mechanistically distinct from nucleosome binding.
Polycomb Group (PcG) proteins are essential regulators of development that maintain gene silencing in Drosophila and mammals through alterations of chromatin structure. One key PcG protein, Posterior Sex Combs (PSC), is part of at least two complexes: Polycomb Repressive Complex 1 (PRC1) and dRING-Associated Factors (dRAF). PRC1-class complexes compact chromatin and inhibit chromatin remodeling, while dRAF has E3 ligase activity for ubiquitylation of histone H2A; activities of both complexes can inhibit transcription. The noncovalent effects of PRC1-class complexes on chromatin can be recapitulated by PSC alone, and the region of PSC required for these activities is essential for PSC function in vivo. To understand how PSC interacts with chromatin to exert its repressive effects, we compared the ability of PSC to bind to and inhibit remodeling of various nucleosomal templates and determined which regions of PSC are required for mononucleosome binding and inhibition of chromatin remodeling. We find that PSC binds mononucleosome templates but inhibits their remodeling poorly. Addition of linker DNA to mononucleosomes allows their remodeling to be inhibited, although higher concentrations of PSC are required than for inhibition of multinucleosome templates. The C-terminal region of PSC (amino acids 456−1603) is important for inhibition of chromatin remodeling, and we identified amino acids 456−909 as being sufficient for stable nucleosome binding but not for inhibition of chromatin remodeling. Our data suggest distinct mechanistic steps between nucleosome binding and inhibition of chromatin remodeling.
Lo
Stanley M
SM
Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA.
Francis
Nicole J
NJ
D000818:Animals; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004330:Drosophila / Q000378:metabolism*; D029721:Drosophila Proteins / Q000378:metabolism*; D006801:Humans; D009707:Nucleosomes / Q000378:metabolism*; D063150:Polycomb Repressive Complex 1; D011485:Protein Binding; D013379:Substrate Specificity
D002843:Chromatin; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D009707:Nucleosomes; C071050:Psc protein, Drosophila; D063150:Polycomb Repressive Complex 1