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Polycomb Group (PcG) proteins are epigenetic repressors essential for control of development and cell differentiation. They form multiple complexes of which PRC1 and PRC2 are evolutionary conserved and obligatory for repression. The targeting of PRC1 and PRC2 is poorly understood and was proposed to be hierarchical and involve tri-methylation of histone H3 (H3K27me3) and/or monoubiquitylation of histone H2A (H2AK118ub). Here, we present a strict test of this hypothesis using the Drosophila model. We discover that neither H3K27me3 nor H2AK118ub is required for targeting PRC complexes to Polycomb Response Elements (PREs). We find that PRC1 can bind PREs in the absence of PRC2 but at many PREs PRC2 requires PRC1 to be targeted. We show that one role of H3K27me3 is to allow PcG complexes anchored at PREs to interact with surrounding chromatin. In contrast, the bulk of H2AK118ub is unrelated to PcG repression. These findings radically change our view of how PcG repression is targeted and suggest that PRC1 and PRC2 can communicate independently of histone modifications., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)

A 4.8 kilobase mouse embryo DNA fragment was inserted into a phage lambda genome and was subsequently characterized by electron microscopy, restriction enzyme mapping and partial DNA sequencing. This fragment contains a 400 base sequence which is homologous to that of an immunoglobulin light lambda chain mRNA which spans 3.3 to 3.7 kilobases from one end of the fragment. Restriction enzyme mapping as well as partial nucleotide sequencing of the 3' terminal of the homology region confirm the previous conclusion [Tonegawa, Brack, Hozumi and Schuller, Proc. Natl. Acad. Sci. USA. 74, 3518-3522 (1977)] that the cloned DNA fragment contains a Vlambda gene sequence which is separate from any Clambda sequence.

The sequence of the entire 5S RNA gene of Drosophila melanogaster was determined by sequencing collectively 23 copies contained in a cloned fragment of Drosophila DNA and by sequencing individually four subcloned gene copies. A repetitive heptamer (GCTG CCT) present in variable numbers immediately following the coding sequence, is responsible for the length heterogeneity in the spacer region. Some of the gene copies contain a nucleotide change in the coding region which results in a new site for the restriction enzyme Mn1 I. The variant 5S RNA produced by these gene copies has not been detected in vivo. Two other single nucleotide variations were identified in the spacer region.

The OR operator region of phage 434 contains three 14 bp blocks with sequence ACAAGA-A--TTGT which are presumed to be the 434 repressor recognition sites. Operator constitutive mutations are located in two of these blocks, while a mutation affecting repressor levels in the lysogenic state is located in the third. Two transcripts obtained in vitro, one leftwards and one rightwards, are tentatively identified as the Prm and PR transcription starts. the arrangement of the 434 operator region appears to be very similar to that of the OR of phage lambda.