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Molecular and Cellular Biology, May 2000, p. 3069-3078, Vol. 20, No. 9
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
A Drosophila ESC-E(Z) Protein Complex Is
Distinct from Other Polycomb Group Complexes and Contains
Covalently Modified ESC
Joyce
Ng,
Craig M.
Hart,
Kelly
Morgan, and
Jeffrey A.
Simon*
Department of Genetics, Cell Biology and
Development and Department of Biochemistry, Molecular Biology and
Biophysics, University of Minnesota, Minneapolis, Minnesota 55455
Received 23 December 1999/Accepted 2 February 2000
The extra sex combs (ESC) and Enhancer of zeste [E(Z)] proteins,
members of the Polycomb group (PcG) of transcriptional repressors, interact directly and are coassociated in fly embryos. We report that
these two proteins are components of a 600-kDa complex in embryos.
Using gel filtration and affinity chromatography, we show that this
complex is biochemically distinct from previously described complexes
containing the PcG proteins Polyhomeotic, Polycomb, and Sex comb on
midleg. In addition, we present evidence that ESC is
phosphorylated in vivo and that this modified ESC is preferentially
associated in the complex with E(Z). Modified ESC accumulates between 2 and 6 h of embryogenesis, which is the developmental time when
esc function is first required. We find that mutations in
E(z) reduce the ratio of modified to unmodified ESC in vivo. We have also generated germ line transformants that express ESC proteins bearing site-directed mutations that disrupt ESC-E(Z) binding in vitro. These mutant ESC proteins fail to provide esc function, show reduced levels of
modification in vivo, and are still assembled into complexes. Taken
together, these results suggest that ESC phosphorylation normally
occurs after assembly into ESC-E(Z) complexes and that it contributes
to the function or regulation of these complexes. We discuss how
biochemically separable ESC-E(Z) and PC-PH complexes might work
together to provide PcG repression.
*
Corresponding author. Mailing address: 321 Church St.
S.E., Minneapolis, MN 55455. Phone: (612) 626-5097. Fax: (612)
626-7031. E-mail: simon{at}biosci.cbs.umn.edu.
Molecular and Cellular Biology, May 2000, p. 3069-3078, Vol. 20, No. 9
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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