CAL1 is the Drosophila CENP-A assembly factor.
Centromeres are specified epigenetically by the incorporation of the histone H3 variant CENP-A. In humans, amphibians, and fungi, CENP-A is deposited at centromeres by the HJURP/Scm3 family of assembly factors, but homologues of these chaperones are absent from a number of major eukaryotic lineages such as insects, fish, nematodes, and plants. In Drosophila, centromeric deposition of CENP-A requires the fly-specific protein CAL1. Here, we show that targeting CAL1 to noncentromeric DNA in Drosophila cells is sufficient to heritably recruit CENP-A, kinetochore proteins, and microtubule attachments. CAL1 selectively interacts with CENP-A and is sufficient to assemble CENP-A nucleosomes that display properties consistent with left-handed octamers. The CENP-A assembly activity of CAL1 resides within an N-terminal domain, whereas the C terminus mediates centromere recognition through an interaction with CENP-C. Collectively, this work identifies the "missing" CENP-A chaperone in flies, revealing fundamental conservation between insect and vertebrate centromere-specification mechanisms.
Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT 06269.
Chin-Chi
CC
Chen
Mekonnen Lemma
ML
Dechassa
Emily
E
Bettini
Mary B
MB
Ledoux
Christian
C
Belisario
Patrick
P
Heun
Karolin
K
Luger
Barbara G
BG
Mellone
D000818:Animals; D001324:Autoantigens / Q000378:metabolism*; D002460:Cell Line; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D020090:Chromosome Segregation; D004278:DNA, Superhelical / Q000378:metabolism; D029721:Drosophila Proteins / Q000737:chemistry / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D044127:Epigenesis, Genetic; D006657:Histones / Q000378:metabolism; D018386:Kinetochores / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism; D011485:Protein Binding; D021382:Protein Sorting Signals
D001324:Autoantigens; C000619700:CENPA protein, human; C558282:Cal1 protein, Drosophila; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D004278:DNA, Superhelical; D029721:Drosophila Proteins; D006657:Histones; D008869:Microtubule-Associated Proteins; C547532:Ndc80 protein, Drosophila; D009707:Nucleosomes; D021382:Protein Sorting Signals; C060905:centromere protein C