{"database":"biostudies-literature","file_versions":[],"scores":{"citationCount":0,"reanalysisCount":0,"viewCount":78,"searchCount":0},"additional":{"submitter":["Barfoot T"],"funding":["NIGMS NIH HHS"],"pagination":["23905-15"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC4583041"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["290(39)"],"pubmed_abstract":["Rad50 and Mre11 form a complex involved in the detection and processing of DNA double strand breaks. Rad50 contains an anti-parallel coiled-coil with two absolutely conserved cysteine residues at its apex. These cysteine residues serve as a dimerization domain and bind a Zn(2+) cation in a tetrathiolate coordination complex known as the zinc-hook. Mutation of the zinc-hook in bacteriophage T4 is lethal, indicating the ability to bind Zn(2+) is critical for the functioning of the MR complex. In vitro, we found that complex formation between Rad50 and a peptide corresponding to the C-terminal domain of Mre11 enhances the ATPase activity of Rad50, supporting the hypothesis that the coiled-coil is a major conduit for communication between Mre11 and Rad50. We constructed mutations to perturb this domain in the bacteriophage T4 Rad50 homolog. Deletion of the Rad50 coiled-coil and zinc-hook eliminates Mre11 binding and ATPase activation but does not affect its basal activity. Mutation of the zinc-hook or disruption of the coiled-coil does not affect Mre11 or DNA binding, but their activation of Rad50 ATPase activity is abolished. Although these mutants excise a single nucleotide at a normal rate, they lack processivity and have reduced repetitive exonuclease rates. Restricting the mobility of the coiled-coil eliminates ATPase activation and repetitive exonuclease activity, but the ability to support single nucleotide excision is retained. These results suggest that the coiled-coiled domain adopts at least two conformations throughout the ATPase/nuclease cycle, with one conformation supporting enhanced ATPase activity and processivity and the other supporting nucleotide excision."],"journal":["The Journal of biological chemistry"],"pubmed_title":["Functional Analysis of the Bacteriophage T4 Rad50 Homolog (gp46) Coiled-coil Domain."],"pmcid":["PMC4583041"],"funding_grant_id":["R01 GM066934"],"pubmed_authors":["Kreuzer KN","Behning BR","Herdendorf TJ","Gao Y","Nelson SW","Stohr BA","Barfoot T"],"view_count":["78"],"additional_accession":[]},"is_claimable":false,"name":"Functional Analysis of the Bacteriophage T4 Rad50 Homolog (gp46) Coiled-coil Domain.","description":"Rad50 and Mre11 form a complex involved in the detection and processing of DNA double strand breaks. Rad50 contains an anti-parallel coiled-coil with two absolutely conserved cysteine residues at its apex. These cysteine residues serve as a dimerization domain and bind a Zn(2+) cation in a tetrathiolate coordination complex known as the zinc-hook. Mutation of the zinc-hook in bacteriophage T4 is lethal, indicating the ability to bind Zn(2+) is critical for the functioning of the MR complex. In vitro, we found that complex formation between Rad50 and a peptide corresponding to the C-terminal domain of Mre11 enhances the ATPase activity of Rad50, supporting the hypothesis that the coiled-coil is a major conduit for communication between Mre11 and Rad50. We constructed mutations to perturb this domain in the bacteriophage T4 Rad50 homolog. Deletion of the Rad50 coiled-coil and zinc-hook eliminates Mre11 binding and ATPase activation but does not affect its basal activity. Mutation of the zinc-hook or disruption of the coiled-coil does not affect Mre11 or DNA binding, but their activation of Rad50 ATPase activity is abolished. Although these mutants excise a single nucleotide at a normal rate, they lack processivity and have reduced repetitive exonuclease rates. Restricting the mobility of the coiled-coil eliminates ATPase activation and repetitive exonuclease activity, but the ability to support single nucleotide excision is retained. These results suggest that the coiled-coiled domain adopts at least two conformations throughout the ATPase/nuclease cycle, with one conformation supporting enhanced ATPase activity and processivity and the other supporting nucleotide excision.","dates":{"release":"2015-01-01T00:00:00Z","publication":"2015 Sep","modification":"2024-11-12T08:56:14.909Z","creation":"2019-03-27T01:58:58Z"},"accession":"S-EPMC4583041","cross_references":{"pubmed":["26242734"],"doi":["10.1074/jbc.m115.675132","10.1074/jbc.M115.675132"]}}