title: CENP-C facilitates the recruitment of M18BP1 to centromeric chromatin.
abstract: Centromeres are important structural constituents of chromosomes that ensure proper chromosome segregation during mitosis by providing defined sites for kinetochore attachment. In higher eukaryotes, centromeres have no specific DNA sequence and thus, they are rather determined through epigenetic mechanisms. A fundamental process in centromere establishment is the incorporation of the histone variant CENP-A into centromeric chromatin, which provides a binding platform for the other centromeric proteins. The Mis18 complex, and, in particular, its member M18BP1 was shown to be essential for both incorporation and maintenance of CENP-A. Here we show that M18BP1 displays a cell cycle-regulated association with centromeric chromatin in mouse embryonic stem cells. M18BP1 is highly enriched at centromeric regions from late anaphase through to G1 phase. An interaction screen against 16 core centromeric proteins revealed a novel interaction of M18BP1 with CENP-C. We mapped the interaction domain in M18BP1 to a central region containing a conserved SANT domain and in CENP-C to the C-terminus. Knock-down of CENP-C leads to reduced M18BP1 association and lower CENP-A levels at centromeres, suggesting that CENP-C works as an important factor for centromeric M18BP1 recruitment and thus for maintaining centromeric CENP-A.
authors.affiliation: Ludwig Maximilians University and Munich Center for Integrated Protein Science (CiPSM), Munich, Germany.
authors.forename: Silvia
authors.initials: S
authors.lastname: Dambacher
authors.forename: Wen
authors.initials: W
authors.lastname: Deng
authors.forename: Matthias
authors.initials: M
authors.lastname: Hahn
authors.forename: Dennis
authors.initials: D
authors.lastname: Sadic
authors.forename: Jonathan
authors.initials: J
authors.lastname: Fröhlich
authors.forename: Alexander
authors.initials: A
authors.lastname: Nuber
authors.forename: Christian
authors.initials: C
authors.lastname: Hoischen
authors.forename: Stephan
authors.initials: S
authors.lastname: Diekmann
authors.forename: Heinrich
authors.initials: H
authors.lastname: Leonhardt
authors.forename: Gunnar
authors.initials: G
authors.lastname: Schotta
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002503:Centromere / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D034622:RNA Interference
chemical_list: D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; C517147:MIS18BP1 protein, human; C060905:centromere protein C