Cloning of a cDNA encoding human centrin, an EF-hand protein of centrosomes and mitotic spindle poles

Ramesh Errabolu, Mark A. Sanders, Jeffrey L Salisbury

Research output: Contribution to journalArticle

156 Citations (Scopus)

Abstract

A human cDNA expression library was screened using anti-centrin antibodies to obtain a cDNA clone encoding human centrin. The cDNA clone contains an open reading frame of 516 base pairs and predicts a product of 172 amino acids with a calculated molecular mass of 19,528 and a pI of 4.61. Sequence analysis demonstrates that human centrin and centrins from higher plants, protozoa, algae, Xenopus and the yeast CDC31 gene product are closely related members of a subfamily of the EF-hand superfamily of calcium-binding proteins. The human centrin sequence has four putative calcium-binding domains as defined by the EF-hand consensus. Immunoprecipitation and western blot studies from HeLa cells confirm that human centrin is a protein of -20,000 M(r) as predicted from the cDNA clone. Indirect immunofluorescence analysis of HeLa cells demonstrates that centrin is localized at the centrosome of interphase cells and that it redistributes to the region of the spindle poles during mitosis. When taken together with earlier genetic studies, these results demonstrate that centrin is a ubiquitous component of centrosomes and mitotic spindle poles of diverse organisms and suggest that centrin plays a role in centrosome separation at the time of mitosis.

Original languageEnglish (US)
Pages (from-to)9-16
Number of pages8
JournalJournal of Cell Science
Volume107
Issue number1
StatePublished - Jan 1994

Fingerprint

Spindle Poles
EF Hand Motifs
Centrosome
Spindle Apparatus
Sulfamethoxazole Drug Combination Trimethoprim
Organism Cloning
Complementary DNA
Proteins
Clone Cells
HeLa Cells
Mitosis
Calcium-Binding Proteins
Interphase
Indirect Fluorescent Antibody Technique
Xenopus
Gene Library
Immunoprecipitation
Base Pairing
Open Reading Frames
Sequence Analysis

Keywords

  • Centrin
  • Centriole
  • Centrosome
  • EF-hand protein
  • Microtubule
  • Mitosis

ASJC Scopus subject areas

  • Cell Biology

Cite this

Cloning of a cDNA encoding human centrin, an EF-hand protein of centrosomes and mitotic spindle poles. / Errabolu, Ramesh; Sanders, Mark A.; Salisbury, Jeffrey L.

In: Journal of Cell Science, Vol. 107, No. 1, 01.1994, p. 9-16.

Research output: Contribution to journalArticle

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