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The catalytic residues of Tn3 resolvase

Olorunniji, FJ and Stark, WM (2009) The catalytic residues of Tn3 resolvase. Nucleic Acids Research, 37 (22). pp. 7590-7602. ISSN 0305-1048

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Abstract

To characterize the residues that participate in the catalysis of DNA cleavage and rejoining by the site-specific recombinase Tn 3 resolvase, we mutated conserved polar or charged residues in the catalytic domain of an activated resolvase variant. We analysed the effects of mutations at 14 residues on proficiency in binding to the recombination site (‘site I’), formation of a synaptic complex between two site Is, DNA cleavage and recombination. Mutations of Y6, R8, S10, D36, R68 and R71 resulted in greatly reduced cleavage and recombination activity, suggesting crucial roles of these six residues in catalysis, whereas mutations of the other residues had less dramatic effects. No mutations strongly inhibited binding of resolvase to site I, but several caused conspicuous changes in the yield or stability of the synapse of two site Is observed by non-denaturing gel electrophoresis. The involvement of some residues in both synapsis and catalysis suggests that they contribute to a regulatory mechanism, in which engagement of catalytic residues with the substrate is coupled to correct assembly of the synapse.

Item Type: Article
Uncontrolled Keywords: 05 Environmental Sciences, 06 Biological Sciences, 08 Information and Computing Sciences
Subjects: R Medicine > RS Pharmacy and materia medica
Divisions: Pharmacy & Biomolecular Sciences
Publisher: Oxford University Press (OUP)
Related URLs:
Date Deposited: 06 Mar 2020 12:01
Last Modified: 06 Mar 2020 12:15
DOI or Identification number: 10.1093/nar/gkp797
URI: http://researchonline.ljmu.ac.uk/id/eprint/12425

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