Functional Evaluation of Bacteriophage T4 Rad50 Signature Motif Residues

dc.contributor.author Herdendorf, Timothy
dc.contributor.author Nelson, Scott
dc.contributor.author Nelson, Scott
dc.contributor.department Biochemistry, Biophysics and Molecular Biology
dc.date 2018-02-17T15:05:52.000
dc.date.accessioned 2020-06-29T23:47:23Z
dc.date.available 2020-06-29T23:47:23Z
dc.date.copyright Sat Jan 01 00:00:00 UTC 2011
dc.date.issued 2011-06-01
dc.description.abstract <p>The repair of DNA double-strand breaks (DSBs) is essential to maintaining the integrity of the genome, and organisms have evolved a conserved mechanism to facilitate their repair. In eukaryotes, archaea, and some bacteriophage, a complex made up of Mre11 and Rad50 (MR complex), which are a nuclease and ATPase, respectively, is involved in the initial processing of DSBs. Rad50 is a member of the ATP Binding Cassette (ABC) protein superfamily, the members of which contain an important Signature motif that acts <em>in trans</em> to complete the dimeric ATP binding site. To explore the functional relevance of this motif, four of its five residues were mutated in bacteriophage T4 Rad50, and their respective ATPase and nuclease activities were evaluated. The mutations reveal the functional roles of the Signature motif in ATP binding, hydrolysis, and cooperativity. In several mutants, the degree of DNA activation of ATP hydrolysis activity is reduced, indicating that the Signature motif is involved in allosteric signal transmission between the DNA and ATP binding sites of the MR complex. ATP hydrolysis is not required for nuclease activity when the probe is near the beginning of the DNA substrate; however, when an internal probe is used, decreases in ATPase activity have substantial effects on nuclease activity, suggesting that ATP hydrolysis is involved in translocation of the complex. Unexpectedly, the ATP hydrolysis and nuclease activities are not directly correlated with each other, and each mutation appears to differentially affect the exonuclease activity of Mre11.</p>
dc.description.comments <p>Reprinted (adapted) with permission from <em>Biochemistry</em> 50 (2011): 6030, doi: <a href="http://dx.doi.org/10.1021/bi200184w" target="_blank">10.1021/bi200184w</a>. Copyright 2011 American Chemical Society.</p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/bbmb_ag_pubs/67/
dc.identifier.articleid 1078
dc.identifier.contextkey 8372522
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath bbmb_ag_pubs/67
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/10800
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/bbmb_ag_pubs/67/0-2011_NelsonSW_FunctionalEvaluationBacteriophage.pdf|||Sat Jan 15 01:27:02 UTC 2022
dc.source.bitstream archive/lib.dr.iastate.edu/bbmb_ag_pubs/67/2011_NelsonSW_FunctionalEvaluationBacteriophage.pdf|||Sat Jan 15 01:27:03 UTC 2022
dc.source.uri 10.1021/bi200184w
dc.subject.disciplines Biochemistry, Biophysics, and Structural Biology
dc.subject.disciplines Molecular Biology
dc.subject.keywords Allosteric signal
dc.subject.keywords Archaea
dc.subject.keywords ATP binding
dc.subject.keywords Double-strand breaks
dc.subject.keywords Bacteriophage T4
dc.subject.keywords DNA Repair
dc.title Functional Evaluation of Bacteriophage T4 Rad50 Signature Motif Residues
dc.type article
dc.type.genre article
dspace.entity.type Publication
relation.isAuthorOfPublication 6570190c-e045-441a-a9bf-59c716840114
relation.isOrgUnitOfPublication c70f85ae-e0cd-4dce-96b5-4388aac08b3f
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