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PMID: 15809444 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Molecular organization of the Ndc80 complex, an essential kinetochore component.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 102 ·No. 15 ·2005-04-12 ·Pages 5363-7

Wei RR, Sorger PK, Harrison SC

Abstract

The four-protein Ndc80 complex, an essential kinetochore component conserved from yeast to humans, plays an indispensable role in proper chromosome alignment and segregation during mitosis. In higher eukaryotes, the homologous complex probably resides in the middle domain of the trilaminar kinetochore, linking centromeric heterochromatin with microtubule-associated structures. We have prepared recombinant Ndc80 complex by pairwise coexpression of its components (Ndc80p and Nuf2p; Spc24p and Spc25p) and shown that they form independently stable subcomplexes. Rotary shadowing electron microscopy, combined with limited proteolysis and antibody labeling, demonstrates that the heterotetrameric Ndc80 complex is an approximately 570-A-long rod, with globular regions at either end. The shaft contains alpha-helical coiled-coil segments from each of the two subcomplexes, linked end-to-end. When integrated with published observations derived from inactivating the components of Ndc80, the molecular organization we deduce suggests that the Spc24p/Spc25p end of the rod faces the centromere and the Ndc80p/Nuf2p end faces a spindle microtubule.

MeSH Terms
Animals Cell Line Centromere/metabolism Chromosomal Proteins, Non-Histone/chemistry,genetics,metabolism,ultrastructure Kinetochores/chemistry Microtubules/metabolism Multiprotein Complexes/chemistry,genetics,metabolism,ultrastructure Nuclear Proteins/chemistry,genetics,metabolism,ultrastructure Protein Conformation Protein Structure, Secondary Protein Subunits/chemistry,genetics,metabolism Saccharomyces cerevisiae/chemistry Saccharomyces cerevisiae Proteins/chemistry,genetics,metabolism,ultrastructure Spodoptera
Chemicals
Chromosomal Proteins, Non-Histone Multiprotein Complexes NDC80 protein, S cerevisiae NUF2 protein, S cerevisiae Nuclear Proteins Protein Subunits Saccharomyces cerevisiae Proteins Spc24 protein, S cerevisiae Spc25 protein, S cerevisiae
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Wei Ronnie R
Department of Biological Chemistry and Molecular Pharmacology, and Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02115, USA.
Sorger Peter K
Harrison Stephen C
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0501168102
Published
2005-04-12
Epub
2005-00-04
Pages
5363-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC555962
Subset
IM
Grants
NIGMS NIH HHS · R01 GM051464 · United States
NIGMS NIH HHS · GM 51464 · United States
External Links
PubMed source DOI: 10.1073/pnas.0501168102 PMC Full Text View journal details
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