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

J Cell Biol. 1993 Dec;123(5):1175-84. doi: 10.1083/jcb.123.5.1175.

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.

Publication types

  • Comparative Study

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*
  • Fungal Proteins / immunology
  • Fungal Proteins / isolation & purification
  • Gene Expression Regulation, Fungal*
  • Genes, Fungal*
  • Microscopy, Electron
  • Molecular Sequence Data
  • Nuclear Proteins / genetics*
  • Nuclear Proteins / immunology
  • Nuclear Proteins / isolation & purification
  • Peptide Fragments / biosynthesis
  • RNA, Messenger / metabolism
  • Recombinant Proteins / biosynthesis
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / ultrastructure
  • Saccharomyces cerevisiae Proteins*
  • Sequence Deletion
  • Sequence Homology, Amino Acid
  • Spindle Apparatus / metabolism*
  • Spindle Apparatus / ultrastructure

Substances

  • Calmodulin-Binding Proteins
  • Cytoskeletal Proteins
  • Epitopes
  • Fungal Proteins
  • Nuclear Proteins
  • Peptide Fragments
  • RNA, Messenger
  • Recombinant Proteins
  • SPC110 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins

Associated data

  • GENBANK/X73297