Further Tests of a Recombination Model in Which {chi} Removes the RecD Subunit From the RecBCD Enzyme of Escherichia coli

Further Tests of a Recombination Model in Which {chi} Removes the RecD Subunit From the RecBCD... F. W. Stahl, L. C. Thomason, I. Siddiqi and M. M. Stahl Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229 When one of two infecting {lambda} phage types in a replication-blocked cross is {chi}(+) and DNA packaging is divorced from the RecBCD-{chi} interaction, complementary {chi}-stimulated recombinants are recovered equally in mass lysates only if the {chi}(+) parent is in excess in the infecting parental mixture. Otherwise, the {chi}(0) recombinant is recovered in excess. This observation implies that, along with the {chi}(0) chromosome, two {chi}(+) parent chromosomes are involved in the formation of each {chi}(+) recombinant. The trimolecular nature of {chi}(+)-stimulated recombination is manifest in recombination between {lambda} and a plasmid. When {lambda} recombines with a plasmid via the RecBCD pathway, the resulting chromosome has an enhanced probability of undergoing {lambda} X {lambda} recombination in the interval into which the plasmid was incorporated. These two observations support a model in which DNA is degraded by Exo V from cos, the sequence that determines the end of packaged {lambda} DNA and acts as point of entry for RecBCD enzyme, to {chi}, the DNA sequence that stimulates the RecBCD enzyme to effect recombination. The model supposes that {chi} acts by ejecting the RecD subunit from the RecBCD enzyme with two consequences. (1) ExoV activity is blocked leaving a highly recombinagenic, frayed duplex end near {chi}, and (2) as the enzyme stripped of the RecD subunit travels beyond {chi} it is competent to catalyze reciprocal recombination. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Genetics Genetics Society of America

Further Tests of a Recombination Model in Which {chi} Removes the RecD Subunit From the RecBCD Enzyme of Escherichia coli

Genetics, Volume 126 (3): 519 – Nov 1, 1990

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Publisher
Genetics Society of America
Copyright
Copyright © 1990 by the Genetics Society of America
ISSN
0016-6731
eISSN
1943-2631
Publisher site
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Abstract

F. W. Stahl, L. C. Thomason, I. Siddiqi and M. M. Stahl Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229 When one of two infecting {lambda} phage types in a replication-blocked cross is {chi}(+) and DNA packaging is divorced from the RecBCD-{chi} interaction, complementary {chi}-stimulated recombinants are recovered equally in mass lysates only if the {chi}(+) parent is in excess in the infecting parental mixture. Otherwise, the {chi}(0) recombinant is recovered in excess. This observation implies that, along with the {chi}(0) chromosome, two {chi}(+) parent chromosomes are involved in the formation of each {chi}(+) recombinant. The trimolecular nature of {chi}(+)-stimulated recombination is manifest in recombination between {lambda} and a plasmid. When {lambda} recombines with a plasmid via the RecBCD pathway, the resulting chromosome has an enhanced probability of undergoing {lambda} X {lambda} recombination in the interval into which the plasmid was incorporated. These two observations support a model in which DNA is degraded by Exo V from cos, the sequence that determines the end of packaged {lambda} DNA and acts as point of entry for RecBCD enzyme, to {chi}, the DNA sequence that stimulates the RecBCD enzyme to effect recombination. The model supposes that {chi} acts by ejecting the RecD subunit from the RecBCD enzyme with two consequences. (1) ExoV activity is blocked leaving a highly recombinagenic, frayed duplex end near {chi}, and (2) as the enzyme stripped of the RecD subunit travels beyond {chi} it is competent to catalyze reciprocal recombination.

Journal

GeneticsGenetics Society of America

Published: Nov 1, 1990

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