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An Adenosine Triphosphate- Dependent 5′-3′ DNA Helicase From sk1-Like Lactococcus lactis F13 Phage

Chmielewska-Jeznach, Magdalena and Steczkiewicz, Kamil and Kobylecki, Kamil and Bardowski, Jacek K. and Szczepankowska, Agnieszka K. (2022) An Adenosine Triphosphate- Dependent 5′-3′ DNA Helicase From sk1-Like Lactococcus lactis F13 Phage. Frontiers in Microbiology, 13 . ISSN 1664-302X

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Official URL: https://www.frontiersin.org/journals/microbiology#

Abstract

Here, we describe functional characterization of an early gene (gp46) product of a virulent Lactococcus lactis sk1-like phage, vB_Llc_bIBBF13 (abbr. F13). The GP46F13 protein carries a catalytically active RecA-like domain belonging to the P-loop NTPase superfamily. It also retains features characteristic for ATPases forming oligomers. In order to elucidate its detailed molecular function, we cloned and overexpressed the gp46 gene in Escherichia coli. Purified GP46F13 protein binds to DNA and exhibits DNA unwinding activity on branched substrates in the presence of adenosine triphosphate (ATP). Size exclusion chromatography with multi-angle light scattering (SEC-MALS) experiments demonstrate that GP46F13 forms oligomers, and further pull-down assays show that GP46F13 interacts with host proteins involved in replication (i.e., DnaK, DnaJ, topoisomerase I, and single-strand binding protein). Taking together the localization of the gene and the obtained results, GP46F13 is the first protein encoded in the early-expressed gene region with helicase activity that has been identified among lytic L. lactis phages up to date.

Item Type:Article
Uncontrolled Keywords:Lactococcus lactis, sk1-like bacteriophage, P-loop NTPase, DNA unwinding, DNA replication
Subjects:Q Science > QH Natural history > QH301 Biology
Q Science > QR Microbiology > QR355 Virology
Divisions:Department of Bioinformatics
Department of Microbial Biochemistry
Laboratory of RNA Biology and Functional Genomics
ID Code:2142
Deposited By: Dr Agnieszka K Szczepankowska
Deposited On:15 Mar 2022 08:16
Last Modified:15 Mar 2022 08:16

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