Cell Cycle-Regulated Attachment of the Ubiquitin-Related Protein Sumo to the Yeast Septins

doi:10.1083/jcb.147.5.981

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© The Rockefeller University Press, 0021-9525/1999//981 $5.00
The Journal of Cell Biology, Volume 147, Number 5, , 1999 981-994

Original Article

Cell Cycle–Regulated Attachment of the Ubiquitin-Related Protein Sumo to the Yeast Septins

Erica S. Johnson^a and Günter Blobel^a

^a Laboratory of Cell Biology, Howard Hughes Medical Institute, Rockefeller University, New York, New York 10021
Department of Biochemistry and Molecular Pharmacology, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107.(215) 503-5393(215) 503-4616

Erica.Johnson{at}mail.tju.edu

SUMO is a ubiquitin-related protein that functions as a posttranslationalmodification on other proteins. SUMO conjugation is essentialfor viability in Saccharomyces cerevisiae and is required forentry into mitosis. We have found that SUMO is attached to theseptins Cdc3, Cdc11, and Shs1/Sep7 specifically during mitosis,with conjugates appearing shortly before anaphase onset anddisappearing abruptly at cytokinesis. Septins are componentsof a belt of 10-nm filaments encircling the yeast bud neck.Intriguingly, only septins on the mother cell side of the budneck are sumoylated. We have identified four major SUMO attachment-sitelysine residues in Cdc3, one in Cdc11, and two in Shs1, allwithin the consensus sequence (IVL)KX(ED). Mutating these siteseliminated the vast majority of bud neck-associated SUMO, aswell as the bulk of total SUMO conjugates in G₂/M-arrested cells,indicating that sumoylated septins are the most abundant SUMOconjugates at this point in the cell cycle. This mutant hasa striking defect in disassembly of septin rings, resultingin accumulation of septin rings marking previous division sites.Thus, SUMO conjugation plays a role in regulating septin ringdynamics during the cell cycle.

Key Words: Smt3 • SUMO • septin • cytoskeleton • protein processing

We are calling the product of the Saccharomyces cerevisiae SMT3gene SUMO to bring the yeast terminology for this modificationin line with the mammalian terminology.

Abbreviations used in this paper: GFP, green fluorescence protein;HA, influenza virus hemagglutinin epitope tag; MALDI-TOF, matrix-assistedlaser desorption/ionization time of flight; NTA, nitriloaceticacid; RanGAP1, Ran GTPase-activating protein; ts, temperaturesensitive; Ub, ubiquitin.

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