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PMID: 8668174 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of overlapping DNA-binding and centromere-targeting domains in the human kinetochore protein CENP-C.

Molecular and cellular biology ·Vol. 16 ·No. 7 ·1996-07-00 ·Pages 3576-86

Yang CH, Tomkiel J, Saitoh H, Johnson DH, Earnshaw WC

Abstract

The kinetochore in eukaryotes serves as the chromosomal site of attachment for microtubules of the mitotic spindle and directs the movements necessary for proper chromosome segregation. In mammalian cells, the kinetochore is a highly differentiated trilaminar structure situated at the surface of the centromeric heterochromatin. CENP-C is a basic, DNA-binding protein that localizes to the inner kinetochore plate, the region that abuts the heterochromatin. Microinjection experiments using antibodies specific for CENP-C have demonstrated that this protein is required for the assembly and/or stability of the kinetochore as well as for a timely transition through mitosis. From these observations, it has been suggested that CENP-C is a structural protein that is involved in the organization or the kinetochore. In this report, we wished to identify and map the functional domains of CENP-C. Analysis of CENP-C truncation mutants expressed in vivo demonstrated that CENP-C possesses an autonomous centromere-targeting domain situated at the central region of the CENP-C polypeptide. Similarly, in vitro assays revealed that a region of CENP-C with the ability to bind DNA is also located at the center of the CENP-C molecule, where it overlaps the centromere-targeting domain.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites Centromere/metabolism Chromosomal Proteins, Non-Histone/biosynthesis,chemistry,metabolism Cloning, Molecular DNA-Binding Proteins/biosynthesis,chemistry,metabolism Gene Expression Humans Mammals Mitosis Molecular Sequence Data Protein Structure, Secondary Recombinant Proteins/biosynthesis,chemistry,metabolism Sequence Deletion
Chemicals
Chromosomal Proteins, Non-Histone DNA-Binding Proteins Recombinant Proteins centromere protein C
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Yang C H
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Tomkiel J
Saitoh H
Johnson D H
Earnshaw W C
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.7.3576
Published
1996-07-00
Pages
3576-86
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231353
Subset
IM
Grants
NIGMS NIH HHS · GM35212 · United States
Databases
GENBANK
U57992, U57993, U57994, U57995, U57996, U57997, U57998
External Links
PubMed source DOI: 10.1128/MCB.16.7.3576 PMC Full Text View journal details
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