Molecular analysis of recombinase-mediated cassette exchange reactions catalyzed by integrase of coliphage HK022

Natalia Malchin; Tatiana Molotsky; Ezra Yagil; Alexander B. Kotlyar; Mikhail Kolot

doi:10.1016/j.resmic.2008.09.005

Molecular analysis of recombinase-mediated cassette exchange reactions catalyzed by integrase of coliphage HK022

Natalia Malchin, Tatiana Molotsky, Ezra Yagil, Alexander B. Kotlyar, Mikhail Kolot^*

^*Corresponding author for this work

Biochemistry Molecular Biology

Research output: Contribution to journal › Article › peer-review

Abstract

The integrase (Int) protein of coliphage HK022 can catalyze in Escherichia coli as well as in in vitro integrative and excisive recombinase-mediated cassette exchange reactions between plasmids as substrates. Atomic force microscopy images have revealed that in the protein-DNA complexes that are formed, the plasmid substrates are connected via one and not two pairs of attachment sites. This observation, together with the elucidation of intermediate co-integrates between the two circular plasmids, suggest that a sequential mechanism of the RMCE reaction is possible.

Original language	English
Pages (from-to)	663-670
Number of pages	8
Journal	Research in Microbiology
Volume	159
Issue number	9-10
DOIs	https://doi.org/10.1016/j.resmic.2008.09.005
State	Published - Nov 2008

Keywords

Atomic force microscopy
Bacteriophage HK022
Integrase
Recombinase-mediated cassette exchange
Site-specific recombination

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10.1016/j.resmic.2008.09.005

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title = "Molecular analysis of recombinase-mediated cassette exchange reactions catalyzed by integrase of coliphage HK022",

abstract = "The integrase (Int) protein of coliphage HK022 can catalyze in Escherichia coli as well as in in vitro integrative and excisive recombinase-mediated cassette exchange reactions between plasmids as substrates. Atomic force microscopy images have revealed that in the protein-DNA complexes that are formed, the plasmid substrates are connected via one and not two pairs of attachment sites. This observation, together with the elucidation of intermediate co-integrates between the two circular plasmids, suggest that a sequential mechanism of the RMCE reaction is possible.",

keywords = "Atomic force microscopy, Bacteriophage HK022, Integrase, Recombinase-mediated cassette exchange, Site-specific recombination",

author = "Natalia Malchin and Tatiana Molotsky and Ezra Yagil and Kotlyar, {Alexander B.} and Mikhail Kolot",

note = "Funding Information: This study was supported by a grant from the US–Israel Binational Science Foundation (grant 2003304) to E.Y. and M.K., by a grant from the Adams Supercenter for Brain Studies to M.K. and by a European grant for Future and Emerging Technologies (FP6-029192) to A.B.K.",

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doi = "10.1016/j.resmic.2008.09.005",

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AU - Kotlyar, Alexander B.

AU - Kolot, Mikhail

N1 - Funding Information: This study was supported by a grant from the US–Israel Binational Science Foundation (grant 2003304) to E.Y. and M.K., by a grant from the Adams Supercenter for Brain Studies to M.K. and by a European grant for Future and Emerging Technologies (FP6-029192) to A.B.K.

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N2 - The integrase (Int) protein of coliphage HK022 can catalyze in Escherichia coli as well as in in vitro integrative and excisive recombinase-mediated cassette exchange reactions between plasmids as substrates. Atomic force microscopy images have revealed that in the protein-DNA complexes that are formed, the plasmid substrates are connected via one and not two pairs of attachment sites. This observation, together with the elucidation of intermediate co-integrates between the two circular plasmids, suggest that a sequential mechanism of the RMCE reaction is possible.

AB - The integrase (Int) protein of coliphage HK022 can catalyze in Escherichia coli as well as in in vitro integrative and excisive recombinase-mediated cassette exchange reactions between plasmids as substrates. Atomic force microscopy images have revealed that in the protein-DNA complexes that are formed, the plasmid substrates are connected via one and not two pairs of attachment sites. This observation, together with the elucidation of intermediate co-integrates between the two circular plasmids, suggest that a sequential mechanism of the RMCE reaction is possible.

KW - Atomic force microscopy

KW - Bacteriophage HK022

KW - Integrase

KW - Recombinase-mediated cassette exchange

KW - Site-specific recombination

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Molecular analysis of recombinase-mediated cassette exchange reactions catalyzed by integrase of coliphage HK022

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