Structural and functional insights into the interaction between the bacteriophage T4 DNA processing proteins gp32 and Dda.

Autor:	He X; Department of Structural Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place MS311, Memphis, TN 38105, USA., Yun MK; Department of Host-Microbe Interactions, St. Jude Children's Research Hospital, 262 Danny Thomas Place MS221, Memphis, TN 38105, USA., Li Z; Department of Structural Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place MS311, Memphis, TN 38105, USA., Waddell MB; Hartwell Center for Biotechnology, St. Jude Children's Research Hospital, 262 Danny Thomas Place MS1300, Memphis, TN 38105, USA., Nourse A; Department of Structural Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place MS311, Memphis, TN 38105, USA., Churion KA; Department of Structural Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place MS311, Memphis, TN 38105, USA., Kreuzer KN; Department of Biochemistry, Duke University Medical Center, Nanaline H. Duke Box 3711, Durham, NC 27710, USA., Byrd AK; Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, 4301 W. Markham Street Slot 516, Little Rock, AR 72205, USA., White SW; Department of Structural Biology, St. Jude Children's Research Hospital, 262 Danny Thomas Place MS311, Memphis, TN 38105, USA.
Jazyk:	angličtina
Zdroj:	Nucleic acids research [Nucleic Acids Res] 2024 Nov 11; Vol. 52 (20), pp. 12748-12762.
DOI:	10.1093/nar/gkae910
Abstrakt:	Bacteriophage T4 is a classic model system for studying the mechanisms of DNA processing. A key protein in T4 DNA processing is the gp32 single-stranded DNA-binding protein. gp32 has two key functions: it binds cooperatively to single-stranded DNA (ssDNA) to protect it from nucleases and remove regions of secondary structure, and it recruits proteins to initiate DNA processes including replication and repair. Dda is a T4 helicase recruited by gp32, and we purified and crystallized a gp32-Dda-ssDNA complex. The low-resolution structure revealed how the C-terminus of gp32 engages Dda. Analytical ultracentrifugation analyses were consistent with the crystal structure. An optimal Dda binding peptide from the gp32 C-terminus was identified using surface plasmon resonance. The crystal structure of the Dda-peptide complex was consistent with the corresponding interaction in the gp32-Dda-ssDNA structure. A Dda-dependent DNA unwinding assay supported the structural conclusions and confirmed that the bound gp32 sequesters the ssDNA generated by Dda. The structure of the gp32-Dda-ssDNA complex, together with the known structure of the gp32 body, reveals the entire ssDNA binding surface of gp32. gp32-Dda-ssDNA complexes in the crystal are connected by the N-terminal region of one gp32 binding to an adjacent gp32, and this provides key insights into this interaction. (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)
Databáze:	MEDLINE
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