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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.


Related in: MedlinePlus

SUMO-dependent extraction of proteins from the chromatin.(A) ssDNA-activated SUMO E3 ligase sumoylates DNA-bound protein and induces its dissociation. (B) Delay in dissociation results in SUMO chain formation through multiple rounds of protein sumoylation. Subsequent ubiqutylation by STUbL promotes Cdc48/Npl4/Ufd1 loading, protein extraction and degradation via proteasome. (C) When the extraction is compromised (e.g., covalent protein–DNA adduct), the protein is processed by Cdc48/Wss1/Doa1 complex. Wss1 is targeted to sumoylated protein via its SIMs and promotes extension of SUMO chain that in return could further stimulate Wss1 accumulation and oligomerization at the site of DNA damage (Wss1 foci). Binding to ssDNA and oligomerization triggers metalloprotease activity of Wss1 and initiates substrate processing. The process is assisted by Cdc48 and Doa1 and finally ends in the vacuole.DOI:http://dx.doi.org/10.7554/eLife.06763.033
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fig11: SUMO-dependent extraction of proteins from the chromatin.(A) ssDNA-activated SUMO E3 ligase sumoylates DNA-bound protein and induces its dissociation. (B) Delay in dissociation results in SUMO chain formation through multiple rounds of protein sumoylation. Subsequent ubiqutylation by STUbL promotes Cdc48/Npl4/Ufd1 loading, protein extraction and degradation via proteasome. (C) When the extraction is compromised (e.g., covalent protein–DNA adduct), the protein is processed by Cdc48/Wss1/Doa1 complex. Wss1 is targeted to sumoylated protein via its SIMs and promotes extension of SUMO chain that in return could further stimulate Wss1 accumulation and oligomerization at the site of DNA damage (Wss1 foci). Binding to ssDNA and oligomerization triggers metalloprotease activity of Wss1 and initiates substrate processing. The process is assisted by Cdc48 and Doa1 and finally ends in the vacuole.DOI:http://dx.doi.org/10.7554/eLife.06763.033

Mentions: Our data suggest a model for the SUMO-dependent extraction of proteins from chromatin (Figure 11). DNA lesions induce exposure of ssDNA and sumoylation of nearby chromatin-bound proteins (Psakhye and Jentsch, 2012). Thus, exposure of ssDNA and/or initial sumoylation of proteins at the sites of DNA damage could provide binding sites for Wss1. Its intrinsic ligase activity could then sumoylate other proteins, extending SUMO chains, become proteolytically activated. Wss1 would also recruit Doa1/Cdc48 to disassemble proteins from the damage site(s) and target the complexes to the vacuole. The proteolytic activation of Wss1 both down regulates Wss1 mediated action and fragments other proteins to induce their dissociation. Thus, we propose that autophagy processes poor proteasome substrates formed as a result of DNA damage, that is, protein–protein and protein–nucleic acid crosslinks, irreversibly trapped enzymes and abnormal nucleic acid intermediates (Figure 11). Alternatively, or in addition, STUBL could be recruited to ubiquitinate chromosomal lesions that have not been repaired and recruit Doa1/Ufd1/Npl4 to chaperone ubiquitinated substrates to the proteasome.10.7554/eLife.06763.033Figure 11.SUMO-dependent extraction of proteins from the chromatin.


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)

SUMO-dependent extraction of proteins from the chromatin.(A) ssDNA-activated SUMO E3 ligase sumoylates DNA-bound protein and induces its dissociation. (B) Delay in dissociation results in SUMO chain formation through multiple rounds of protein sumoylation. Subsequent ubiqutylation by STUbL promotes Cdc48/Npl4/Ufd1 loading, protein extraction and degradation via proteasome. (C) When the extraction is compromised (e.g., covalent protein–DNA adduct), the protein is processed by Cdc48/Wss1/Doa1 complex. Wss1 is targeted to sumoylated protein via its SIMs and promotes extension of SUMO chain that in return could further stimulate Wss1 accumulation and oligomerization at the site of DNA damage (Wss1 foci). Binding to ssDNA and oligomerization triggers metalloprotease activity of Wss1 and initiates substrate processing. The process is assisted by Cdc48 and Doa1 and finally ends in the vacuole.DOI:http://dx.doi.org/10.7554/eLife.06763.033
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4559962&req=5

fig11: SUMO-dependent extraction of proteins from the chromatin.(A) ssDNA-activated SUMO E3 ligase sumoylates DNA-bound protein and induces its dissociation. (B) Delay in dissociation results in SUMO chain formation through multiple rounds of protein sumoylation. Subsequent ubiqutylation by STUbL promotes Cdc48/Npl4/Ufd1 loading, protein extraction and degradation via proteasome. (C) When the extraction is compromised (e.g., covalent protein–DNA adduct), the protein is processed by Cdc48/Wss1/Doa1 complex. Wss1 is targeted to sumoylated protein via its SIMs and promotes extension of SUMO chain that in return could further stimulate Wss1 accumulation and oligomerization at the site of DNA damage (Wss1 foci). Binding to ssDNA and oligomerization triggers metalloprotease activity of Wss1 and initiates substrate processing. The process is assisted by Cdc48 and Doa1 and finally ends in the vacuole.DOI:http://dx.doi.org/10.7554/eLife.06763.033
Mentions: Our data suggest a model for the SUMO-dependent extraction of proteins from chromatin (Figure 11). DNA lesions induce exposure of ssDNA and sumoylation of nearby chromatin-bound proteins (Psakhye and Jentsch, 2012). Thus, exposure of ssDNA and/or initial sumoylation of proteins at the sites of DNA damage could provide binding sites for Wss1. Its intrinsic ligase activity could then sumoylate other proteins, extending SUMO chains, become proteolytically activated. Wss1 would also recruit Doa1/Cdc48 to disassemble proteins from the damage site(s) and target the complexes to the vacuole. The proteolytic activation of Wss1 both down regulates Wss1 mediated action and fragments other proteins to induce their dissociation. Thus, we propose that autophagy processes poor proteasome substrates formed as a result of DNA damage, that is, protein–protein and protein–nucleic acid crosslinks, irreversibly trapped enzymes and abnormal nucleic acid intermediates (Figure 11). Alternatively, or in addition, STUBL could be recruited to ubiquitinate chromosomal lesions that have not been repaired and recruit Doa1/Ufd1/Npl4 to chaperone ubiquitinated substrates to the proteasome.10.7554/eLife.06763.033Figure 11.SUMO-dependent extraction of proteins from the chromatin.

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