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Protein-oligonucleotide conjugates as model substrates for DNA-protein crosslink repair proteases.

ABSTRACT: Covalent DNA-protein crosslinks (DPCs) have emerged as pervasive sources of genome instability. DPCs are targeted for repair by DNA-dependent proteases of the Wss1/SPRTN family. However, understanding how these enzymes achieve specificity has been hampered by the lack of suitable in vitro model substrates. Here, we describe the generation of defined protein-oligonucleotide conjugates as DPC model substrates, which enable the analysis of DPC proteases in activity and binding assays. For complete details on the use and execution of this protocol, please refer to Reinking et al. (2020).

SUBMITTER: Reinking HK

PROVIDER: S-EPMC8219891 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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Protein-oligonucleotide conjugates as model substrates for DNA-protein crosslink repair proteases.

Reinking Hannah K HK Stingele Julian J

STAR protocols 20210615 2

Covalent DNA-protein crosslinks (DPCs) have emerged as pervasive sources of genome instability. DPCs are targeted for repair by DNA-dependent proteases of the Wss1/SPRTN family. However, understanding how these enzymes achieve specificity has been hampered by the lack of suitable <i>in vitro</i> model substrates. Here, we describe the generation of defined protein-oligonucleotide conjugates as DPC model substrates, which enable the analysis of DPC proteases in activity and binding assays. For co ...[more]

PMID: 34189469

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