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Structural and functional characterization of TraI from pKM101 reveals basis for DNA processing

View ORCID ProfileAnnika Breidenstein, View ORCID ProfileJosy ter Beek, View ORCID ProfileRonnie P-A Berntsson
doi: https://doi.org/10.1101/2022.06.02.494495
Annika Breidenstein
1Department of Medical Biochemistry and Biophysics, Umeå University, SE-90187 Umeå, Sweden
2Wallenberg Centre for Molecular Medicine, Umeå University, Umeå, Sweden
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Josy ter Beek
1Department of Medical Biochemistry and Biophysics, Umeå University, SE-90187 Umeå, Sweden
2Wallenberg Centre for Molecular Medicine, Umeå University, Umeå, Sweden
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  • For correspondence: josy.beek@umu.se ronnie.berntsson@umu.se
Ronnie P-A Berntsson
1Department of Medical Biochemistry and Biophysics, Umeå University, SE-90187 Umeå, Sweden
2Wallenberg Centre for Molecular Medicine, Umeå University, Umeå, Sweden
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  • For correspondence: josy.beek@umu.se ronnie.berntsson@umu.se
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Abstract

To support conjugation via Type 4 Secretion Systems, relaxases perform the initial processing of the substrate DNA. TraI from the well-studied conjugative plasmid pKM101 is one such relaxase, which has not yet been characterized. Here, we report the crystal structure of the trans-esterase domain of TraI in complex with its substrate oriT DNA, highlighting the conserved DNA binding mechanism. Additionally, we present an apo structure of the trans-esterase domain of TraI, which allows us to visualize the conformational changes involved in DNA binding. We also show a complete characterization of the DNA binding, nicking and religation of the trans-esterase domain, helicase domain and full-length TraI. Furthermore, TraI is shown to be a tetramer. These results reveal that the trans-esterase domain behaves in a similar way as its homologs TraI from the F plasmid and TrwC from R388, but the tetramerization of the full-length protein highlights a significant difference that affects the function of TraI.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 02, 2022.
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Structural and functional characterization of TraI from pKM101 reveals basis for DNA processing
Annika Breidenstein, Josy ter Beek, Ronnie P-A Berntsson
bioRxiv 2022.06.02.494495; doi: https://doi.org/10.1101/2022.06.02.494495
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Structural and functional characterization of TraI from pKM101 reveals basis for DNA processing
Annika Breidenstein, Josy ter Beek, Ronnie P-A Berntsson
bioRxiv 2022.06.02.494495; doi: https://doi.org/10.1101/2022.06.02.494495

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