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PMID: 15133482 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Sgt1 is required for human kinetochore assembly.

EMBO reports ·Vol. 5 ·No. 6 ·2004-06-00 ·Pages 626-31

Steensgaard P, Garrè M, Muradore I, Transidico P, Nigg EA, Kitagawa K, Earnshaw WC, Faretta M, Musacchio A

Abstract

Budding yeast Sgt1 is required for kinetochore assembly, and its homologues have a role in cAMP signalling in fungi and pathogen resistance in plants. The function of mammalian Sgt1 is unknown. We report that RNA interference-mediated depletion of Sgt1 from HeLa cells causes dramatic alterations of the mitotic spindle and problems in chromosome alignment. Cells lacking Sgt1 undergo a mitotic delay due to activation of the spindle checkpoint. The checkpoint response, however, is significantly weakened in Sgt1-depleted cells, and this correlates with a dramatic reduction in kinetochore levels of Mad1, Mad2 and BubR1. These effects are explained by a problem in kinetochore assembly that prevents the localization of Hec1, CENP-E, CENP-F, CENP-I, but not CENP-C, to mitotic kinetochores. Our studies implicate Sgt1 as an essential protein and a critical assembly factor for the mammalian kinetochore, and lend credit to the hypothesis of a kinetochore assembly pathway that is conserved from yeast to man.

MeSH Terms
Calcium-Binding Proteins/analysis,immunology Cell Cycle Proteins/genetics,physiology Centromere/immunology,metabolism Chromosomal Proteins, Non-Histone/analysis,immunology DNA-Binding Proteins/analysis,immunology HeLa Cells Humans Kinetochores/metabolism,ultrastructure Mad2 Proteins Mitosis/genetics,physiology Nuclear Proteins Phosphoproteins/analysis,immunology Protein Kinases/analysis,immunology Protein Serine-Threonine Kinases RNA Interference RNA, Small Interfering/metabolism Repressor Proteins/analysis,immunology Spindle Apparatus/genetics,metabolism
Chemicals
CENPI protein, human Calcium-Binding Proteins Cell Cycle Proteins Chromosomal Proteins, Non-Histone DNA-Binding Proteins MAD1L1 protein, human MAD2L1 protein, human Mad2 Proteins Nuclear Proteins Phosphoproteins RNA, Small Interfering Repressor Proteins SUGT1 protein, human Protein Kinases BUB1 protein, human Bub1 spindle checkpoint protein Protein Serine-Threonine Kinases
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Steensgaard Peter
Department of Experimental Oncology, European Institute of Oncology, Via Ripamonti 435, 20141 Milan, Italy.
Garrè Massimiliano
Muradore Ivan
Transidico Pietro
Nigg Erich A
Kitagawa Katsumi
Earnshaw William C
Faretta Mario
Musacchio Andrea
References (32)
32 references, click to expand
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Article Info
Journal
EMBO reports
Abbr.
EMBO Rep
ISSN
1469-221X
DOI
10.1038/sj.embor.7400154
Published
2004-06-00
Epub
2004-00-07
Pages
626-31
Language
English
Region
England
NLM ID
100963049
PMCID
PMC1299074
Subset
IM
Grants
NIGMS NIH HHS · GM68418 · United States
NCI NIH HHS · CA21765 · United States
NCI NIH HHS · P30 CA021765 · United States
NIGMS NIH HHS · R01 GM068418 · United States
Wellcome Trust · 073915 · United Kingdom
External Links
PubMed source DOI: 10.1038/sj.embor.7400154 PMC Full Text View journal details
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