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PDBsum entry 2wkc
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DNA binding protein
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PDB id
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2wkc
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References listed in PDB file
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Key reference
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Title
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Structure and function of phage p2 orf34(p2), A new type of single-Stranded DNA binding protein.
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Authors
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E.Scaltriti,
M.Tegoni,
C.Rivetti,
H.Launay,
J.Y.Masson,
A.H.Magadan,
D.Tremblay,
S.Moineau,
R.Ramoni,
J.Lichière,
V.Campanacci,
C.Cambillau,
M.Ortiz-Lombardía.
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Ref.
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Mol Microbiol, 2009,
73,
1156-1170.
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PubMed id
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Abstract
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Lactococcus lactis, a Gram-positive bacterium widely used by the dairy industry,
is subject to infection by a diverse population of virulent phages,
predominantly by those of the 936 group, including the siphovirus phage p2.
Confronted with the negative impact of phage infection on milk fermentation, the
study of the biology of lactococcal provides insight from applied and
fundamental perspectives. We decided to characterize the product of the orf34
gene from lactococcus phage p2, which was considered as a candidate
single-stranded DNA binding protein (SSB) due to its localization downstream of
a gene coding for a single-strand annealing protein. Two-dimensional gel
electrophoresis showed that ORF34(p2) is expressed in large amounts during the
early phases of phage infection, suggesting an important role in this process.
Gel-shift assays, surface plasmon resonance and atomic force microscopy
demonstrated that ORF34(p2) interacts with single-strand DNA with nanomolar
affinity. We also determined the crystal structure of ORF34(p2) and showed that
it bears a variation of the typical oligonucleotide/oligosaccharide binding-fold
of SSBs. Finally, we found that ORF34(p2) is able to stimulate Escherichia coli
RecA-mediated homologous recombination. The specific structural and biochemical
properties that distinguish ORF34(p2) from other SSB proteins are discussed.
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