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PMID: 7503995 Published · ppublish English Comparative Study Journal Article

A spacer protein in the Saccharomyces cerevisiae spindle poly body whose transcript is cell cycle-regulated.

The Journal of cell biology ·Vol. 123 ·No. 5 ·1993-12-00 ·Pages 1175-84

Kilmartin JV, Dyos SL, Kershaw D, Finch JT

Abstract

Monoclonal antibodies against the 110-kD component of the yeast spindle pole body (SPB) were used to clone the corresponding gene SPC110. SPC110 is identical to NUF1 (Mirzayan, C., C. S. Copeland, and M. Synder. 1992. J. Cell Biol. 116:1319-1332). SPC110/NUF1 has an MluI cell cycle box consensus sequence in its putative promoter region, and we found that the transcript was cell cycle regulated in a similar way to other MluI-regulated transcripts. Spc110p/Nuflp has a long central region with a predicted coiled-coil structure. We expressed this region in Escherichia coli and showed by rotary shadowing that rods of the predicted length were present. The 110-kD component is localized in the SPB to the gap between the central plaque and the sealed ends of the nuclear microtubules near the inner plaque (Rout, M., and J. V. Kilmartin. 1990. J. Cell Biol. 111:1913-1927). We found that rodlike structures bridge this gap. When truncations of SPC110 with deletions in the coiled-coil region of the protein replaced the wild-type gene, the gap between the central plaque and the ends of the microtubules decreased in proportion to the size of the deletion. This suggests that Spc110p connects these two parts of the SPB together and that the coiled-coil domain acts as a spacer element.

MeSH Terms
Amino Acid Sequence Base Sequence Calmodulin-Binding Proteins Cell Cycle/physiology Cloning, Molecular Cytoskeletal Proteins DNA Mutational Analysis Epitopes Fluorescent Antibody Technique Fungal Proteins/genetics,immunology,isolation & purification Gene Expression Regulation, Fungal Genes, Fungal Microscopy, Electron Molecular Sequence Data Nuclear Proteins/genetics,immunology,isolation & purification Peptide Fragments/biosynthesis RNA, Messenger/metabolism Recombinant Proteins/biosynthesis Saccharomyces cerevisiae/genetics,metabolism,ultrastructure Saccharomyces cerevisiae Proteins Sequence Deletion Sequence Homology, Amino Acid Spindle Apparatus/metabolism,ultrastructure
Chemicals
Calmodulin-Binding Proteins Cytoskeletal Proteins Epitopes Fungal Proteins Nuclear Proteins Peptide Fragments RNA, Messenger Recombinant Proteins SPC110 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kilmartin J V
MRC Laboratory of Molecular Biology, Cambridge, England.
Dyos S L
Kershaw D
Finch J T
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1993-12-00
Pages
1175-84
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2119877
Subset
IM
Databases
GENBANK
X73297
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