A proposed role for the Polycomb group protein dRING in meiotic sister-chromatid cohesion.
ORD protein is required for accurate chromosome segregation during male and female meiosis in Drosophila melanogaster. Null ord mutations result in random segregation of sister chromatids during both meiotic divisions because cohesion is completely abolished prior to kinetochore capture of microtubules during meiosis I. Previous analyses of mutant ord alleles have led us to propose that the C-terminal half of the ORD protein mediates protein-protein interactions that are essential for sister-chromatid cohesion. To identify proteins that interact with ORD, we conducted a yeast two-hybrid screen using an ORD bait and isolated dRING, a core subunit of the Drosophila Polycomb repressive complex 1. We show that a missense mutation in ORD completely ablates the two-hybrid interaction with dRING and prevents nuclear retention of the mutant ORD protein in male meiotic cells. Using affinity-purified antibodies generated against full-length recombinant dRING, we demonstrate that dRING protein is expressed in the male and female gonads and colocalizes extensively with ORD on the chromatin of primary spermatocytes during G2 of meiosis. Our results suggest a novel role for the Polycomb group protein dRING and are consistent with the model that interaction of dRING and ORD is required to promote the proper segregation of meiotic chromosomes.
Balicky
Eric M
EM
Department of Biological Sciences, Dartmouth College, Hanover, NH 03755, USA.
Young
Lynn
L
Orr-Weaver
Terry L
TL
Bickel
Sharon E
SE
D000818:Animals; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D002842:Chromatids / Q000235:genetics / Q000378:metabolism*; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D023902:Chromosome Pairing / Q000235:genetics / Q000502:physiology*; D020090:Chromosome Segregation / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D017344:Genes, Insect; D008297:Male; D008540:Meiosis / Q000235:genetics / Q000502:physiology*; D020125:Mutation, Missense / Q000235:genetics; D063150:Polycomb Repressive Complex 1; D011485:Protein Binding / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D013090:Spermatocytes / Q000166:cytology / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D002843:Chromatin; D029721:Drosophila Proteins; C066770:Pc protein, Drosophila; D011994:Recombinant Proteins; D063150:Polycomb Repressive Complex 1; C570575:Sce protein, Drosophila