In vivo functional dissection of human inner kinetochore protein CENP-C.
CENP-C is a fundamental component of the inner kinetochore plate and contributes to the formation of functional centromeres in eukaryotic organisms. Recruitment of CENP-C to kinetochore requires other centromere proteins, particularly CENP-A, CENP-H, and CENP-I. However, how CENP-C is correctly localized at the kinetochore is not clearly determined, mainly due to the functional variety of its domains, which hints at a complex recruitment mechanism. Here, by both immunofluorescent labeling and chromatin/immunoprecipitation we could show that human CENP-C contains two distinct domains, one in the central region, between amino acids 426 and 537, and the second one in the carboxyl terminal region, between amino acids 638 and 943, which are both capable of localizing at centromeres and binding alpha-satellite DNA. The presence of two domains that iterate the same function despite being significantly different in their amino acid sequence and structure suggests that CENP-C may target the centromere by establishing multiple contacts with both the DNA and protein constituents of the kinetochore.
Trazzi
Stefania
S
Department of Biology, University of Bologna, via Selmi 3, Bologna, Italy.
Bernardoni
Roberto
R
Diolaiti
Daniel
D
Politi
Valeria
V
Earnshaw
William C
WC
Perini
Giovanni
G
Della Valle
Giuliano
G
D015153:Blotting, Western; D002503:Centromere / Q000737:chemistry; D002843:Chromatin / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004276:DNA, Satellite / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005557:Formaldehyde / Q000494:pharmacology; D006801:Humans; D018386:Kinetochores / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009154:Mutation; D011233:Precipitin Tests; D017434:Protein Structure, Tertiary
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D003432:Cross-Linking Reagents; D004276:DNA, Satellite; C060905:centromere protein C; D005557:Formaldehyde