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Published April 1989 | Published
Journal Article Open

The Saccharomyces cerevisiae SEC14 Gene Encodes a Cytosolic Factor That Is Required for Transport of Secretory Proteins from the Yeast Golgi Complex


We have obtained and characterized a genomic clone of SEC14, a Saccharomyces cerevisiae gene whose product is required for export of yeast secretory proteins from the Golgi complex. Gene disruption experiments indicated that SEC14 is an essential gene for yeast vegetative growth. Nucleotide sequence analysis revealed the presence of an intron within the SEC14 structural gene, and predicted the synthesis of a hydrophilic polypeptide of 35 kD in molecular mass. In confirmation, immunoprecipitation experiments demonstrated SEC14p to be an unglycosylated polypeptide, with an apparent molecular mass of some 37 kD, that behaved predominantly as a cytosolic protein in subcellular fractionation experiments. These data were consistent with the notion that SEC14p is a cytosolic factor that promotes protein export from yeast Golgi. Additional radiolabeling experiments also revealed the presence of SEC14p-related polypeptides in extracts prepared from the yeasts Kluyveromyces lactis and Schizosaccharomyces pombe. Furthermore, the K. lactis SEC14p was able to functionally complement S. cerevisiae sec14ts defects. These data suggested a degree of conservation of SEC14p structure and function in these yeasts species.

Additional Information

© 1989 Rockefeller University Press. After the Initial Publication Period, RUP will grant to the public the non-exclusive right to copy, distribute, or display the Article under a Creative Commons Attribution-Noncommercial-Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/legalcode, or updates thereof. Received for publication 9 November 1988 and in revised form 15 December 1988. We wish to thank Bill Bedzyk for expert assistance in the raising of SEC 14p antiserum; Tom Stevens, Joel Rothman, and Dan Klionsky for generous gifts of antisera and plasmids; Kurt Eakle for helpful discussions concerning fractionation methods; and Randy Schekman, Eric Phizicky, Jo Anne Wise, and Stephen Johnston for yeast strains. We are particularly grateful to Randy Schekman for helpful discussions throughout the work, and to John Crnnan for critical comments on the manuscript. Special thanks are also extended to Angela Cox for expert preparation of the manuscript. These studies were supported by a grant from the National Science Foundation (DCB-8620076) and the Arnold Beckman Scholar Award to V. A. Bankaitis.

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