Native and recombinant polycomb group complexes establish a selective block to template accessibility to repress transcription in vitro.
Polycomb group (PcG) proteins are responsible for stable repression of homeotic gene expression during Drosophila melanogaster development. They are thought to stabilize chromatin structure to prevent transcription, though how they do this is unknown. We have established an in vitro system in which the PcG complex PRC1 and a recombinant PRC1 core complex (PCC) containing only PcG proteins are able to repress transcription by both RNA polymerase II and by T7 RNA polymerase. We find that assembly of the template into nucleosomes enhances repression by PRC1 and PCC. The subunit Psc is able to inhibit transcription on its own. PRC1- and PCC-repressed templates remain accessible to Gal4-VP16 binding, and incubation of the template with HeLa nuclear extract before the addition of PCC eliminates PCC repression. These results suggest that PcG proteins do not merely prohibit all transcription machinery from binding the template but instead likely inhibit specific steps in the transcription reaction.
King
Ian F G
IF
Department of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Francis
Nicole J
NJ
Kingston
Robert E
RE
D000818:Animals; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D046911:Macromolecular Substances; D009707:Nucleosomes / Q000378:metabolism; D063150:Polycomb Repressive Complex 1; D012319:RNA Polymerase II / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism*; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014764:Viral Proteins
D029721:Drosophila Proteins; D046911:Macromolecular Substances; D009707:Nucleosomes; C066770:Pc protein, Drosophila; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014764:Viral Proteins; D063150:Polycomb Repressive Complex 1; D012319:RNA Polymerase II; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases