Pokhilko, Alexandra, Zhao, Jia, Ebenhoeh, Oliver, Smith, Margaret C. M., Stark, W. Marshall and Colloms, Sean D. (2016). The mechanism of phi C31 integrase directionality: experimental analysis and computational modelling. Nucleic Acids Res., 44 (15). S. 7360 - 7373. OXFORD: OXFORD UNIV PRESS. ISSN 1362-4962

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Abstract

Serine integrases, DNA site-specific recombinases used by bacteriophages for integration and excision of their DNA to and from their host genomes, are increasingly being used as tools for programmed rearrangements of DNA molecules for biotechnology and synthetic biology. A useful feature of serine integrases is the simple regulation and unidirectionality of their reactions. Recombination between the phage attP and host attB sites is promoted by the serine integrase alone, giving recombinant attL and attR sites, whereas the 'reverse' reaction (between attL and attR) requires an additional protein, the recombination directionality factor (RDF). Here, we present new experimental data on the kinetics and regulation of recombination reactions mediated by phi C31 integrase and its RDF, and use these data as the basis for amathematical model of the reactions. The model accounts for the unidirectionality of the attP xattB and attL x attR reactions by hypothesizing the formation of structurally distinct, kinetically stable integrase-DNA product complexes, dependent on the presence or absence of RDF. The model accounts for all the available experimental data, and predicts how mutations of the proteins or alterations of reaction conditions might increase the conversion efficiency of recombination.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Pokhilko, AlexandraUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Zhao, JiaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Ebenhoeh, OliverUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Smith, Margaret C. M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Stark, W. MarshallUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Colloms, Sean D.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-262591
DOI: 10.1093/nar/gkw616
Journal or Publication Title: Nucleic Acids Res.
Volume: 44
Number: 15
Page Range: S. 7360 - 7373
Date: 2016
Publisher: OXFORD UNIV PRESS
Place of Publication: OXFORD
ISSN: 1362-4962
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
SITE-SPECIFIC RECOMBINATION; SERINE RECOMBINASE; DNA; CELLS; INSERTION; SYNAPSIS; BINDING; PROTEIN; SYSTEM; DOMAINMultiple languages
Biochemistry & Molecular BiologyMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/26259

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