Genome-wide analysis reveals a cell cycle-dependent mechanism controlling centromere propagation.
Centromeres are the structural and functional foundation for kinetochore formation, spindle attachment, and chromosome segregation. In this study, we isolated factors required for centromere propagation using genome-wide RNA interference screening for defects in centromere protein A (CENP-A; centromere identifier [CID]) localization in Drosophila melanogaster. We identified the proteins CAL1 and CENP-C as essential factors for CID assembly at the centromere. CID, CAL1, and CENP-C coimmunoprecipitate and are mutually dependent for centromere localization and function. We also identified the mitotic cyclin A (CYCA) and the anaphase-promoting complex (APC) inhibitor RCA1/Emi1 as regulators of centromere propagation. We show that CYCA is centromere localized and that CYCA and RCA1/Emi1 couple centromere assembly to the cell cycle through regulation of the fizzy-related/CDH1 subunit of the APC. Our findings identify essential components of the epigenetic machinery that ensures proper specification and propagation of the centromere and suggest a mechanism for coordinating centromere inheritance with cell division.
Erhardt
Sylvia
S
Department of Genome Dynamics, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Mellone
Barbara G
BG
Betts
Craig M
CM
Zhang
Weiguo
W
Karpen
Gary H
GH
Straight
Aaron F
AF
D064173:Anaphase-Promoting Complex-Cyclosome; D000818:Animals; D017306:Annexin A2 / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D020090:Chromosome Segregation; D019925:Cyclin A / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D044127:Epigenesis, Genetic; D055106:Genome-Wide Association Study; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D008938:Mitosis / Q000235:genetics; D009154:Mutation; D034622:RNA Interference; D009418:S100 Proteins / Q000378:metabolism; D043743:Ubiquitin-Protein Ligase Complexes / Q000378:metabolism; D000099248:S100 Calcium Binding Protein A10
D064173:Anaphase-Promoting Complex-Cyclosome; D017306:Annexin A2; D018797:Cell Cycle Proteins; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D019925:Cyclin A; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D006657:Histones; D009418:S100 Proteins; D043743:Ubiquitin-Protein Ligase Complexes; D000099248:S100 Calcium Binding Protein A10; C427237:Cid protein, Drosophila; C491136:Rca1 protein, Drosophila; C060905:centromere protein C