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PMID: 11557811 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Creation and characterization of temperature-sensitive CENP-C mutants in vertebrate cells.

Nucleic acids research ·Vol. 29 ·No. 18 ·2001-09-15 ·Pages 3796-803

Fukagawa T, Regnier V, Ikemura T

Abstract

CENP-C is an evolutionarily conserved centromere protein that is thought to be an important component in kinetochore assembly in vertebrate cells. However, the functional role of CENP-C in cell cycle progression remains unclear. To further understand CENP-C function, we developed a method incorporating the hyper-recombinogenic chicken B lymphocyte cell line DT40 to create several temperature-sensitive CENP-C mutants in DT40 cells. We found that, under restrictive conditions, one temperature-sensitive mutant, ts4-11, displayed metaphase delay and chromosome missegregation but proceeded through the cell cycle until arrest at G(1) phase. Furthermore, ts4-11 cells were transfected with a human HeLa cell cDNA library maintained in a retroviral vector, and genes that suppressed the temperature-sensitive phenotype were identified. One of these suppressor genes encodes SUMO-1, which is a ubiquitin-like protein. This finding suggests that SUMO-1 may be involved in centromere function in vertebrate cells. The novel strategy reported here will be useful and applicable to a wide range of proteins that have general cell-autonomous function in vertebrate cells.

MeSH Terms
Animals Base Sequence Cell Division/genetics,physiology Cell Survival/genetics,physiology Centromere/metabolism Chickens Chromosomal Proteins, Non-Histone/genetics,physiology Humans Mutation Phenotype Plasmids/genetics SUMO-1 Protein Suppression, Genetic Temperature Time Factors Transfection Ubiquitins/genetics,physiology
Chemicals
Chromosomal Proteins, Non-Histone SUMO-1 Protein Ubiquitins centromere protein C
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Fukagawa T
National Institute of Genetics and Graduate University for Advanced Studies, Mishima, Shizuoka 411-8540, Japan. [email protected]
Regnier V
Ikemura T
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/29.18.3796
Email
[email protected]
Published
2001-09-15
Pages
3796-803
Language
English
Region
England
NLM ID
0411011
PMCID
PMC55920
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/29.18.3796 PMC Full Text View journal details
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