JCB logo
PeproTech: Cell Culture Supplements
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 541K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by VanRheenen, S. M.
Right arrow Articles by Gerard Waters, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by VanRheenen, S. M.
Right arrow Articles by Gerard Waters, M.
Right arrowPubmed/NCBI databases
*Gene*GEO Profiles
*HomoloGene*Protein
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

© The Rockefeller University Press, 0021-9525/1998//1107 $5.00
The Journal of Cell Biology, Volume 141, Number 5, , 1998 1107-1119

Articles

Sec35p, a Novel Peripheral Membrane Protein, Is Required for ER to Golgi Vesicle Docking



Susan M. VanRheenen*, Xiaochun Cao{ddagger}, Vladimir V. Lupashin*, Charles Barlowe{ddagger}, and M. Gerard Waters*

* Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544; and {ddagger} Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755

SEC35 was identified in a novel screen for temperature-sensitive mutants in the secretory pathway of the yeast Saccharomyces cerevisiae (Wuestehube et al., 1996. Genetics. 142:393–406). At the restrictive temperature, the sec35-1 strain exhibits a transport block between the ER and the Golgi apparatus and accumulates numerous vesicles. SEC35 encodes a novel cytosolic protein of 32 kD, peripherally associated with membranes. The temperature-sensitive phenotype of sec35-1 is efficiently suppressed by YPT1, which encodes the rab-like GTPase required early in the secretory pathway, or by SLY1-20, which encodes a dominant form of the ER to Golgi target -SNARE–associated protein Sly1p. Weaker suppression is evident upon overexpression of genes encoding the vesicle-SNAREs SEC22, BET1, or YKT6. The cold-sensitive lethality that results from deleting SEC35 is suppressed by YPT1 or SLY1-20. These genetic relationships suggest that Sec35p acts upstream of, or in conjunction with, Ypt1p and Sly1p as was previously found for Uso1p. Using a cell-free assay that measures distinct steps in vesicle transport from the ER to the Golgi, we find Sec35p is required for a vesicle docking stage catalyzed by Uso1p. These genetic and biochemical results suggest Sec35p acts with Uso1p to dock ER-derived vesicles to the Golgi complex.

Abbreviations used in this paper: CEN, centromere; COP, coat protein; CPY, carboxypeptidase Y; gp-{alpha}-factor, glycosylated pro-{alpha}-factor; GST, glutathione-S-transferase; NSF, N-ethylmaleimide–sensitive fusion protein; ORF, open reading frame; PGK, phosphoglycerate kinase; SC, synthetic complete medium; SNAP, soluble NSF–attachment protein; t-SNARE, target membrane–SNAP receptor; v-SNARE, vesicle-SNAP receptor; YPD, rich medium.

We are very grateful to R. Schekman (University of California, Berkeley, CA) for the sec35 strain. We thank S. Emr, S. Ferro-Novick, J. Gerst, C. Kaiser, P. Novick, H. Pelham, M. Rose (Princeton University, Princeton, NJ), R. Schekman, H.D. Schmitt and members of their laboratories for generously supplying reagents and strains, and D. Hasara (Princeton University) for expert assistance in antibody production. We are particularly grateful to S. Harris, K. Paul, and S. Sapperstein (all three from Princeton University) for critical reading of the manuscript and to all members of the laboratories for helpful discussion.

Address all correspondence to M. Gerard Waters, Department of Molecular Biology, Princeton University, Princeton, NJ 08544. Tel.: (609) 258-2891. Fax: (609) 258-1701. E-mail: gwaters{at}molbio.princeton.edu


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents