esc, E(z), Rpd3 and Caf1 copurify in a complex with apparent molecular mass of about 600 kDa.

Source was embryonic nuclear extract of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

Nuclear extract was fractionated by several chromatographic steps, tracking the distribution of esc- and E(z)-containing fractions by western blot. These fractions were then subjected to anti-FLAG immunoprecipitation and component proteins identified by mass spectroscopy. Identity of copurifying proteins confirmed by western blot.

The association of Caf1 with esc appears to be distinct from other Caf1-containing complexes, since components of other Caf1-containing complexes do not coimmunoprecipitate with esc (for example, Iswi and Caf1-105).

Less than 5% of Caf1 present in embryonic nuclear extracts is present in the esc/E(z)-containing complex.

Source was embryonic nuclear extract of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

Source was embryonic nuclear extract wild-type fly line; proteins produced from endogenous genes.

The complex of Su(z)12, E(z), Rpd3, esc and Caf1 has an apparent size of 600-650 kDa (gel filtration).

A histone methyltransferase activity was purified from embryonic nuclear extracts.

esc, E(z), Su(z)12 and Caf1 copurify as part of a polycomb repressive (PRC2) complex.

Source was baculovirus-Sf9 cell system; bait produced from tagged transgenic construct; prey produced from transgenic construct.

Source was embryonic nuclear extract of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

esc, E(z), Su(z)12 and Caf1 copurify as part of a polycomb repressive (PRC2) complex.

Nuclear extract was fractionated on a QAE-Sepharose comlum, then used for FLAG-affinity purification of esc containing complexes.

Interaction in vitro; bait and prey produced recombinant fusion proteins in baculovirus and insect cell system.

Affinity purification of the esc-E(z) complex was carried out from a mixture of esc, E(z), Su(z)12, and Caf1-55 recombinant proteins. Deletion of the VEFS domain of Su(z)12 results in loss of Caf1-55 from the complex. Deletion of E(z) II domain results in loss of Su(z)12 and Caf1-55 from the complex.

Su(z)12, E(z), Caf1 and esc were coexpressed in Sf9 cells and purified as a complex.

Su(z)12, E(z), Caf1 and esc form the PRC2 (Polycomb repressive complex 2).

Source was baculovirus-infected Sf9 cell extract; proteins produced from transfected constructs.

Source was cell extract of baculovirus-infected Sf9 cell line; bait produced from transfected construct; prey produced from transfected construct.

PRC2 complex of Su(z)12, E(z), esc and Caf1-55.

FLAG-tagged esc was co-expressed with components of the PRC2 complex (Su(z)12, E(z) and Caf1-55) as well as Scm.

Source was cell extract of baculovirus-infected Sf9 cell line; bait produced from transfected construct; prey produced from transfected construct.

Interaction in vitro; bait and prey produced recombinant fusion proteins in baculovirus and insect cell system.

The PRC2 complex consists of esc, E(z) and Caf1-55.