Recognition of oriT for DNA processing at termination of a round of conjugal transfer

Conjugal transfer of plasmid DNA is terminated when the transferred strand, linearized at the 38 base-pair origin of transfer (oriT), is recircularized. For the plasmid R1162, it is the protein MobA, covalently linked to the linear strand, that rejoins the ends by a reversible transesterification re...

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Bibliographic Details
Published in:Journal of molecular biology 2000-07, Vol.300 (5), p.1067-1077
Main Authors: Becker, Eric C, Meyer, Richard J
Format: Article
Language:English
Subjects:
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Base Sequence
Binding, Competitive
Cloning, Molecular
conjugation
Conjugation, Genetic - genetics
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
DNA, Bacterial - metabolism
DNA, Recombinant - genetics
DNA, Single-Stranded - chemistry
DNA, Single-Stranded - genetics
DNA, Single-Stranded - metabolism
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Escherichia coli - genetics
Mutation
Nucleic Acid Conformation
Oligodeoxyribonucleotides - genetics
oriT
plasmid
Plasmids - chemistry
Plasmids - genetics
Plasmids - metabolism
Protein Binding
protein-DNA binding
Regulatory Sequences, Nucleic Acid - genetics
relaxase
Sequence Analysis, DNA
Trans-Activators - genetics
Trans-Activators - metabolism
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Summary:Conjugal transfer of plasmid DNA is terminated when the transferred strand, linearized at the 38 base-pair origin of transfer (oriT), is recircularized. For the plasmid R1162, it is the protein MobA, covalently linked to the linear strand, that rejoins the ends by a reversible transesterification reaction. We have identified from those oligonucleotides with a partially degenerate oriT base sequence, subpopulations bound by MobA that undergo transesterification, and support efficient termination of conjugal transfer. Two domains of oriT, a ten base-pair inverted repeat and an adjacent TAA, are required for tight binding by the protein, whereas the location of the dinucleotide YG determines the site of strand cleavage. The results indicate that capture of MobA by oriT, and subsequent processing of the DNA for termination, are determined by different sequence motifs within this locus.
ISSN:0022-2836
1089-8638
DOI:10.1006/jmbi.2000.3902