In vivo characterization of the nonessential budding yeast anaphase-promoting complex/cyclosome components Swm1p, Mnd2p and Apc9p.
We have examined the in vivo requirement of two recently identified nonessential components of the budding yeast anaphase-promoting complex, Swm1p and Mnd2p, as well as that of the previously identified subunit Apc9p. swm1Delta mutants exhibit synthetic lethality or conditional synthetic lethality with other APC/C subunits and regulators, whereas mnd2Delta mutants are less sensitive to perturbation of the APC/C. swm1Delta mutants, but not mnd2Delta mutants, exhibit defects in APC/C substrate turnover, both during the mitotic cell cycle and in alpha-factor-arrested cells. In contrast, apc9Delta mutants exhibit only minor defects in substrate degradation in alpha-factor-arrested cells. In cycling cells, degradation of Clb2p, but not Pds1p or Clb5p, is delayed in apc9Delta. Our findings suggest that Swm1p is required for full catalytic activity of the APC/C, whereas the requirement of Mnd2p for APC/C function appears to be negligible under standard laboratory conditions. Furthermore, the role of Apc9p in APC/C-dependent ubiquitination may be limited to the proteolysis of a select number of substrates.
Page
Andrew M
AM
Program in Biochemistry, Cellular, and Molecular Biology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Aneliunas
Vicky
V
Lamb
John R
JR
Hieter
Philip
P
D064173:Anaphase-Promoting Complex-Cyclosome; D015153:Blotting, Western; D019926:Cyclin B / Q000378:metabolism; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005690:Galactose / Q000378:metabolism; D005838:Genotype; D047468:Immunoprecipitation; D009154:Mutation / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D004718:Saccharomycetales / Q000235:genetics*; D043743:Ubiquitin-Protein Ligase Complexes / Q000235:genetics*; D044767:Ubiquitin-Protein Ligases; D014452:Ubiquitins / Q000378:metabolism
C093749:CLB2 protein, S cerevisiae; D019926:Cyclin B; C577468:MND2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014452:Ubiquitins; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome; D044767:Ubiquitin-Protein Ligases; C577465:SWM1 protein, S cerevisiae; D005690:Galactose