Chen, Yei-Wei, Su, Bo-Yu, Van Duyne, Gregory D et al. (2 more authors) (2023) The influence of coiled-coil motif of serine recombinase toward the directionality regulation. Biophysical Journal. ISSN 0006-3495
Abstract
Serine integrases promote the recombination of two complementary DNA sequences, attP and attB, to create hybrid sequences, attL and attR. The reaction is unidirectional in the absence of an accessory protein called recombination directionality factor. We utilized tethered particle motion (TPM) experiments to investigate the reaction behaviors of two model serine integrases from Listeria innocua phage LI and Streptomyces coelicolor phage C31. Detailed kinetic analyses of wild-type and mutant proteins were carried out to verify the mechanisms of recombination directionality. In particular, we assessed the influence of a coiled-coil motif (CC) that is conserved in the C-terminal domain of serine integrases and is an important prerequisite for efficient recombination. Compared to wild type, we found that CC deletions in both serine integrases reduced the overall abundance of integrase (Int) att-site complexes and favored the formation of nonproductive complexes over recombination-competent complexes. Furthermore, the rate at which CC mutants formed productive synaptic complexes and disassembled aberrant nonproductive complexes was significantly reduced. It is notable that while the φC31 Int CC is essential for recombination, the LI Int CC plays an auxiliary role for recombination to stabilize protein-protein interactions and to control the directionality of the reaction.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2023 Biophysical Society. |
Keywords: | Phage,Integrases,Synthetic biology,Mechanism,LISTERIA-MONOCYTOGENES,Streptomyces coelicolor,biotechnology |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Biology (York) |
Depositing User: | Pure (York) |
Date Deposited: | 01 Dec 2023 12:00 |
Last Modified: | 17 Dec 2024 00:27 |
Published Version: | https://doi.org/10.1016/j.bpj.2023.11.009 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1016/j.bpj.2023.11.009 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:206106 |
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Filename: PIIS0006349523007099.pdf
Description: The influence of coiled-coil motif of serine recombinase toward the directionality regulation
Licence: CC-BY 2.5