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Wss1 metalloprotease partners with Cdc48/Doa1 in processing genotoxic SUMO conjugates.

Balakirev MY, Mullally JE, Favier A, Assard N, Sulpice E, Lindsey DF, Rulina AV, Gidrol X, Wilkinson KD - Elife (2015)

Bottom Line: Activation of Wss1 results in metalloprotease self-cleavage and proteolysis of associated proteins.In cells lacking Tdp1, clearance of topoisomerase covalent complexes becomes SUMO and Wss1-dependent.Upon genotoxic stress, Wss1 is vacuolar, suggesting a link between genotoxic stress and autophagy involving the Doa1 adapter.

View Article: PubMed Central - PubMed

Affiliation: Institut de recherches en technologies et sciences pour le vivant-Biologie à Grande Echelle, Commissariat a l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.

ABSTRACT
Sumoylation during genotoxic stress regulates the composition of DNA repair complexes. The yeast metalloprotease Wss1 clears chromatin-bound sumoylated proteins. Wss1 and its mammalian analog, DVC1/Spartan, belong to minigluzincins family of proteases. Wss1 proteolytic activity is regulated by a cysteine switch mechanism activated by chemical stress and/or DNA binding. Wss1 is required for cell survival following UV irradiation, the smt3-331 mutation and Camptothecin-induced formation of covalent topoisomerase 1 complexes (Top1cc). Wss1 forms a SUMO-specific ternary complex with the AAA ATPase Cdc48 and an adaptor, Doa1. Upon DNA damage Wss1/Cdc48/Doa1 is recruited to sumoylated targets and catalyzes SUMO chain extension through a newly recognized SUMO ligase activity. Activation of Wss1 results in metalloprotease self-cleavage and proteolysis of associated proteins. In cells lacking Tdp1, clearance of topoisomerase covalent complexes becomes SUMO and Wss1-dependent. Upon genotoxic stress, Wss1 is vacuolar, suggesting a link between genotoxic stress and autophagy involving the Doa1 adapter.

No MeSH data available.


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Structural organization of WLM proteins.(A) Wss1 domain organization was determined by BLOCKS program by analyzing 12 WLM proteins. (B) Conserved motifs shared by WLM proteins are shown. Amino acid residues potentially involved in the organization of the active site and in the regulation of enzymatic activity are marked by asterisk. (C) Structural comparison of WLM and SprT families using Wss1 and DVC1/Spartan as examples. DVC1/Spartan and 2 proteins from WLM and SprT families with similar domain organization are shown.DOI:http://dx.doi.org/10.7554/eLife.06763.004
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fig1s1: Structural organization of WLM proteins.(A) Wss1 domain organization was determined by BLOCKS program by analyzing 12 WLM proteins. (B) Conserved motifs shared by WLM proteins are shown. Amino acid residues potentially involved in the organization of the active site and in the regulation of enzymatic activity are marked by asterisk. (C) Structural comparison of WLM and SprT families using Wss1 and DVC1/Spartan as examples. DVC1/Spartan and 2 proteins from WLM and SprT families with similar domain organization are shown.DOI:http://dx.doi.org/10.7554/eLife.06763.004

Mentions: Wss1 is predicted to be a metalloprotease, the prototype of the WLM superfamily of proteins (Wss1-like metalloproteases) (Iyer et al., 2004). Sequence alignment of Wss1 with 11 related proteins revealed highly conserved motifs, the most extensive being the metalloprotease WLM domain (Figure 1A and Figure 1—figure supplement 1). Consensus secondary structure prediction and 3D modeling support a metzincin-like fold for the WLM domain (Gomis-Ruth, 2003, Figure 1—figure supplement 2) most similar to minigluzincins (Lopez-Pelegrin et al., 2013), and the closest mammalian Wss1 analog, DVC1/Spartan (Figure 1—figure supplement 2). Additional motifs are two Cdc48-binding sequences (SHP and VIM), and a putative SUMO-interacting motif, SIM2 (Figure 1A). Another SIM, SIM1, is located just upstream of SIM2 and is also present in all 12 proteins. Finally, these modeling results suggest that the N-terminal region of Wss1 might adopt a beta-grasp fold characteristic of SUMO and Ub (data not shown).10.7554/eLife.06763.003Figure 1.Wss1 is a SUMO-targeted metalloprotease.


Wss1 metalloprotease partners with Cdc48/Doa1 in processing genotoxic SUMO conjugates.

Balakirev MY, Mullally JE, Favier A, Assard N, Sulpice E, Lindsey DF, Rulina AV, Gidrol X, Wilkinson KD - Elife (2015)

Structural organization of WLM proteins.(A) Wss1 domain organization was determined by BLOCKS program by analyzing 12 WLM proteins. (B) Conserved motifs shared by WLM proteins are shown. Amino acid residues potentially involved in the organization of the active site and in the regulation of enzymatic activity are marked by asterisk. (C) Structural comparison of WLM and SprT families using Wss1 and DVC1/Spartan as examples. DVC1/Spartan and 2 proteins from WLM and SprT families with similar domain organization are shown.DOI:http://dx.doi.org/10.7554/eLife.06763.004
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4559962&req=5

fig1s1: Structural organization of WLM proteins.(A) Wss1 domain organization was determined by BLOCKS program by analyzing 12 WLM proteins. (B) Conserved motifs shared by WLM proteins are shown. Amino acid residues potentially involved in the organization of the active site and in the regulation of enzymatic activity are marked by asterisk. (C) Structural comparison of WLM and SprT families using Wss1 and DVC1/Spartan as examples. DVC1/Spartan and 2 proteins from WLM and SprT families with similar domain organization are shown.DOI:http://dx.doi.org/10.7554/eLife.06763.004
Mentions: Wss1 is predicted to be a metalloprotease, the prototype of the WLM superfamily of proteins (Wss1-like metalloproteases) (Iyer et al., 2004). Sequence alignment of Wss1 with 11 related proteins revealed highly conserved motifs, the most extensive being the metalloprotease WLM domain (Figure 1A and Figure 1—figure supplement 1). Consensus secondary structure prediction and 3D modeling support a metzincin-like fold for the WLM domain (Gomis-Ruth, 2003, Figure 1—figure supplement 2) most similar to minigluzincins (Lopez-Pelegrin et al., 2013), and the closest mammalian Wss1 analog, DVC1/Spartan (Figure 1—figure supplement 2). Additional motifs are two Cdc48-binding sequences (SHP and VIM), and a putative SUMO-interacting motif, SIM2 (Figure 1A). Another SIM, SIM1, is located just upstream of SIM2 and is also present in all 12 proteins. Finally, these modeling results suggest that the N-terminal region of Wss1 might adopt a beta-grasp fold characteristic of SUMO and Ub (data not shown).10.7554/eLife.06763.003Figure 1.Wss1 is a SUMO-targeted metalloprotease.

Bottom Line: Activation of Wss1 results in metalloprotease self-cleavage and proteolysis of associated proteins.In cells lacking Tdp1, clearance of topoisomerase covalent complexes becomes SUMO and Wss1-dependent.Upon genotoxic stress, Wss1 is vacuolar, suggesting a link between genotoxic stress and autophagy involving the Doa1 adapter.

View Article: PubMed Central - PubMed

Affiliation: Institut de recherches en technologies et sciences pour le vivant-Biologie à Grande Echelle, Commissariat a l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.

ABSTRACT
Sumoylation during genotoxic stress regulates the composition of DNA repair complexes. The yeast metalloprotease Wss1 clears chromatin-bound sumoylated proteins. Wss1 and its mammalian analog, DVC1/Spartan, belong to minigluzincins family of proteases. Wss1 proteolytic activity is regulated by a cysteine switch mechanism activated by chemical stress and/or DNA binding. Wss1 is required for cell survival following UV irradiation, the smt3-331 mutation and Camptothecin-induced formation of covalent topoisomerase 1 complexes (Top1cc). Wss1 forms a SUMO-specific ternary complex with the AAA ATPase Cdc48 and an adaptor, Doa1. Upon DNA damage Wss1/Cdc48/Doa1 is recruited to sumoylated targets and catalyzes SUMO chain extension through a newly recognized SUMO ligase activity. Activation of Wss1 results in metalloprotease self-cleavage and proteolysis of associated proteins. In cells lacking Tdp1, clearance of topoisomerase covalent complexes becomes SUMO and Wss1-dependent. Upon genotoxic stress, Wss1 is vacuolar, suggesting a link between genotoxic stress and autophagy involving the Doa1 adapter.

No MeSH data available.


Related in: MedlinePlus