Xenopus CENP-A assembly into chromatin requires base excision repair proteins.
CENP-A is an essential histone H3 variant found in all eukaryotes examined to date. To begin to determine how CENP-A is assembled into chromatin, we developed a binding assay using sperm chromatin in cell-free extract derived from Xenopus eggs. Our data suggest that the catalytic activities of an unidentified deoxycytidine deaminase and UNG2, a uracil DNA glycosylase, are involved in CENP-A assembly. In support of this model, inhibiting deoxycytidine deaminase with zebularine, or uracil DNA glycosylase with Ugi, uracil or UTP results in a lack of detectable CENP-A on sperm DNA. Conversely, inducing DNA damage increases the level of CENP-A detected on sperm chromatin. Our data suggest that base excision repair may be involved in assembly of this histone H3 variant.
Zeitlin
Samantha G
SG
Department of Biological Sciences, University of California San Diego, Mail Code 0322, La Jolla, CA 92093, USA. szeitlin@biomail.ucsd.edu
Patel
Sheetal
S
Kavli
Bodil
B
Slupphaug
Geir
G
D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D002384:Catalysis; D002467:Cell Nucleus / Q000737:chemistry; D000076247:Centromere Protein A; D002843:Chromatin / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D003564:Cytidine Deaminase; D004249:DNA Damage; D045647:DNA Glycosylases / Q000032:analysis / Q000378:metabolism*; D004260:DNA Repair / Q000502:physiology*; D004261:DNA Replication / Q000187:drug effects; D005260:Female; D016195:G2 Phase; D008297:Male; D008969:Molecular Sequence Data; D009700:Nucleoside Deaminases / Q000378:metabolism*; D010063:Ovum / Q000737:chemistry / Q000378:metabolism; D013094:Spermatozoa / Q000737:chemistry / Q000378:metabolism; D014498:Uracil / Q000032:analysis / Q000378:metabolism; D051981:Uracil-DNA Glycosidase; D029867:Xenopus Proteins / Q000378:metabolism*; D014982:Xenopus laevis
D001324:Autoantigens; D000076247:Centromere Protein A; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D029867:Xenopus Proteins; D014498:Uracil; D045647:DNA Glycosylases; D051981:Uracil-DNA Glycosidase; D009700:Nucleoside Deaminases; D003564:Cytidine Deaminase; C011530:deoxycytidine deaminase