Mlp-dependent anchorage and stabilization of a desumoylating enzyme is required to prevent clonal lethality.
Myosin-like proteins 1 and 2 (Mlp1 and Mlp2) form filaments attached to the nucleoplasmic side of the nuclear pore complexes via interaction with the nucleoporin Nup60. Here, we show that Mlps and Nup60, but not several other nucleoporins, are required to localize and stabilize a desumoylating enzyme Ulp1. Moreover, like Mlps, Ulp1 exhibits a unique asymmetric distribution on the nuclear envelope. Consistent with a role in regulating Ulp1, removal of either or both MLPs affects the SUMO conjugate pattern. We also show that deleting MLPs or the localization domains of Ulp1 results in DNA damage sensitivity and clonal lethality, the latter of which is caused by increased levels of 2-micron circle DNA. Epistatic and dosage suppression analyses further demonstrate that Mlps function upstream of Ulp1 in 2-micron circle maintenance and the damage response. Together, our results reveal that Mlps play important roles in regulating Ulp1 and subsequently affect sumoylation stasis, growth, and DNA repair.
Zhao
Xiaolan
X
Laboratory of Cell Biology, The Rockefeller University, New York, NY 10021, USA.
Wu
Chia-Yung
CY
Blobel
Günter
G
D002999:Clone Cells / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D004270:DNA, Circular / Q000235:genetics; D004795:Enzyme Stability / Q000235:genetics; D004843:Epistasis, Genetic; D018628:Gene Dosage; D005804:Genes, Lethal / Q000235:genetics; D022022:Nuclear Pore / Q000235:genetics / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D016601:RNA-Binding Proteins; D025842:SUMO-1 Protein / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D004270:DNA, Circular; C080988:MLP1 protein, S cerevisiae; C119200:MLP2 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; C495208:Nup60 protein, S cerevisiae; D016601:RNA-Binding Proteins; D025842:SUMO-1 Protein; D029701:Saccharomyces cerevisiae Proteins; D003546:Cysteine Endopeptidases; C118083:Ulp1 protease