PMID:8598302: Difference between revisions

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<span style = 'font-size: large'> '''Blakely, G and Sherratt, D'''  (1996) Determinants of selectivity in Xer site-specific recombination.''Genes Dev.'' '''10''':762-73</span>
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'''Blakely, G and Sherratt, D'''  (1996) Determinants of selectivity in Xer site-specific recombination. ''Genes Dev.'' '''10''':762-73
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!align=left  |Abstract
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A remarkable property of some DNA-binding proteins that can interact with and pair distant DNA segments is that they mediate their biological function only when their binding sites are arranged in a specific configuration. Xer site-specific recombination at natural plasmid recombination sites (e.g., cer in ColE1) is preferentially intramolecular, converting dimers to monomers. In contrast, Xer recombination at the Escherichia coli chromosomal site dif can occur intermolecularly and intramolecularly. Recombination at both types of site requires the cooperative interactions of two related recombinases, XerC and XerD, with a 30-bp recombination core site. The dif core site is sufficient for recombination when XerC and XerD are present, whereas recombination at plasmid sites requires approximately 200 bp of adjacent accessory sequences and accessory proteins. These accessory factors ensure that recombination is intramolecular. Here we use a model system to show that selectivity for intramolecular recombination, and the consequent requirement for accessory factors, can arise by increasing the spacing between XerC- and XerD-binding sites from 6 to 8 bp. This reduces the affinity of the recombinases for the core site and changes the geometry of the recombinase/DNA complex. These changes are correlated with altered interactions of the recombinases with the core site and a reduced efficiency of XerC-mediated cleavage. We propose that the accessory sequences and proteins compensate for these changes and provide a nucleoprotein structure of fixed geometry that can only form and function effectively on circular molecules containing directly repeated sites.
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[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=8598302 PubMed]


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|{{TableHeadingColor}} |'''Abstract''' || <protect>A remarkable property of some DNA-binding proteins that can interact with and pair distant DNA segments is that they mediate their biological function only when their binding sites are arranged in a specific configuration. Xer site-specific recombination at natural plasmid recombination sites (e.g., cer in ColE1) is preferentially intramolecular, converting dimers to monomers. In contrast, Xer recombination at the Escherichia coli chromosomal site dif can occur intermolecularly and intramolecularly. Recombination at both types of site requires the cooperative interactions of two related recombinases, XerC and XerD, with a 30-bp recombination core site. The dif core site is sufficient for recombination when XerC and XerD are present, whereas recombination at plasmid sites requires approximately 200 bp of adjacent accessory sequences and accessory proteins. These accessory factors ensure that recombination is intramolecular. Here we use a model system to show that selectivity for intramolecular recombination, and the consequent requirement for accessory factors, can arise by increasing the spacing between XerC- and XerD-binding sites from 6 to 8 bp. This reduces the affinity of the recombinases for the core site and changes the geometry of the recombinase/DNA complex. These changes are correlated with altered interactions of the recombinases with the core site and a reduced efficiency of XerC-mediated cleavage. We propose that the accessory sequences and proteins compensate for these changes and provide a nucleoprotein structure of fixed geometry that can only form and function effectively on circular molecules containing directly repeated sites.</protect>
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|{{TableHeadingColor}} nowrap |'''Links''' ||  [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=8598302 PubMed]
Amino Acid Sequence; Bacterial Proteins/metabolism; Base Sequence; Binding Sites; Chromosomes, Bacterial/genetics; DNA Footprinting; DNA Nucleotidyltransferases/genetics; DNA Nucleotidyltransferases/metabolism; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; DNA, Superhelical/genetics; DNA-Binding Proteins/genetics; Escherichia coli/genetics; Escherichia coli Proteins; Hydroxyl Radical; Integrases; Models, Genetic; Molecular Sequence Data; Nucleic Acid Conformation; Oligodeoxyribonucleotides; Phenanthrolines; Plasmids; Recombinases; Recombination, Genetic
 
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Latest revision as of 04:55, 15 September 2013

Citation

Blakely, G and Sherratt, D (1996) Determinants of selectivity in Xer site-specific recombination. Genes Dev. 10:762-73

Abstract

A remarkable property of some DNA-binding proteins that can interact with and pair distant DNA segments is that they mediate their biological function only when their binding sites are arranged in a specific configuration. Xer site-specific recombination at natural plasmid recombination sites (e.g., cer in ColE1) is preferentially intramolecular, converting dimers to monomers. In contrast, Xer recombination at the Escherichia coli chromosomal site dif can occur intermolecularly and intramolecularly. Recombination at both types of site requires the cooperative interactions of two related recombinases, XerC and XerD, with a 30-bp recombination core site. The dif core site is sufficient for recombination when XerC and XerD are present, whereas recombination at plasmid sites requires approximately 200 bp of adjacent accessory sequences and accessory proteins. These accessory factors ensure that recombination is intramolecular. Here we use a model system to show that selectivity for intramolecular recombination, and the consequent requirement for accessory factors, can arise by increasing the spacing between XerC- and XerD-binding sites from 6 to 8 bp. This reduces the affinity of the recombinases for the core site and changes the geometry of the recombinase/DNA complex. These changes are correlated with altered interactions of the recombinases with the core site and a reduced efficiency of XerC-mediated cleavage. We propose that the accessory sequences and proteins compensate for these changes and provide a nucleoprotein structure of fixed geometry that can only form and function effectively on circular molecules containing directly repeated sites.

Links

PubMed

Keywords

Amino Acid Sequence; Bacterial Proteins/metabolism; Base Sequence; Binding Sites; Chromosomes, Bacterial/genetics; DNA Footprinting; DNA Nucleotidyltransferases/genetics; DNA Nucleotidyltransferases/metabolism; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; DNA, Superhelical/genetics; DNA-Binding Proteins/genetics; Escherichia coli/genetics; Escherichia coli Proteins; Hydroxyl Radical; Integrases; Models, Genetic; Molecular Sequence Data; Nucleic Acid Conformation; Oligodeoxyribonucleotides; Phenanthrolines; Plasmids; Recombinases; Recombination, Genetic

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