Molecular characterization of the USO1 gene product which is essential for vesicular transport in Saccharomyces cerevisiae

Biochem Biophys Res Commun. 1994 Apr 15;200(1):647-53. doi: 10.1006/bbrc.1994.1497.

Abstract

We have previously shown that USO1 gene required in the protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus encodes a hydrophilic protein of 1790 amino acids. The sequence of carboxyl-terminal 1010 amino acids was predicted to have an alpha-helical structure characteristic of the coiled-coil rod region of the cytoskeleton-related proteins. Antibodies raised against partial sequences of the Uso1 polypeptide reacted with a 200 kDa protein in Western blots of the wild-type yeast proteins. The Uso1 protein was found predominantly in the soluble fraction and displayed a molecular mass of 800-900 kDa in gel filtration when globular protein were used as molecular mass standards. In sucrose density gradient centrifugation, however, the Uso1 protein cosedimented with a globular 6S marker protein, horseradish peroxidase (44 kDa). These results suggest that, in its native state, the Uso1 protein forms a nonglobular oligomer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Carrier Proteins*
  • Centrifugation, Density Gradient
  • Chromatography, Gel
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • Electrophoresis, Polyacrylamide Gel
  • Endoplasmic Reticulum / metabolism*
  • Fungal Proteins / biosynthesis*
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism
  • Genes, Fungal*
  • Golgi Apparatus / metabolism*
  • Molecular Weight
  • Open Reading Frames
  • Rabbits / immunology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins*
  • Vesicular Transport Proteins*

Substances

  • Carrier Proteins
  • DNA, Fungal
  • Fungal Proteins
  • Saccharomyces cerevisiae Proteins
  • USO1 protein, S cerevisiae
  • Vesicular Transport Proteins