Recombination-dependent DNA replication in bacteriophage T4: An evolutionary study.

Persistent Link:
http://hdl.handle.net/10150/185267
Title:
Recombination-dependent DNA replication in bacteriophage T4: An evolutionary study.
Author:
McCreary, Ronald Patrick.
Issue Date:
1990
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Genetic recombination is an ubiquitious process in living organisms and is one of the most elaborate activities that DNA undergoes. As such, the recombination process potentially interacts with all of the major metabolic events involving DNA. This study examined one of these interactions, specifically that occuring between recombination and DNA replication in phage T4. This interaction was investigated from both the mechanisitic and evolutionary viewpoints. In phage and bacteria, the central reactions of homologous recombination are those of homologous pairing and strand insertion. These reactions are generally catalyzed by a single enzyme. The E. coli recA gene product serves as the paradigm for this class of enzyme. Four cloned wild type "recA" genes from the bacterial species E. coli, A. caviae, B. pertussis, and the uvsX gene from phage T4, were used to test for complementation of both E. coli recA⁻ and phage T4 uvsX⁻ mutants. The B. pertussis recA⁺ gene was able to complement an E. coli recA⁻ mutant with respect to the repair of UV induced DNA damage and general recombination. In a phage Mode 2 DNA synthesis assay, all three bacterial recA genes complemented a uvsX⁻ mutant to some extent, but the B. pertussis RecA protein allowed "runaway" DNA synthesis to occur. In contrast, none of the three bacterial genes were able to increase DNA synthesis of a uvsX⁺, uvsY⁻ phage T4. In a one step growth experiment, only the B. pertussis RecA protein complemented a phage uvsX⁻ mutant. However, none of the three bacterial recA genes complemented the same phage mutant in a UV survival assay. These results imply that normal wild type Mode 2 phage DNA synthesis is an example of a specific interaction between the DNA replication and homologous recombination "machines" of phage T4. Multiplicity reactivation (MR) is a type of recombinational repair. MR experiments were performed using a gene 42(ts) mutant phage to study the effect of this member of the phage's DNA replication machinery on recombinational DNA repair. The data show that MR is dependent on the gene 42 gene product suggesting that the phage DNA replication and recombination metabolic pathways are coupled through a protein-protein interaction involving gpuvsX and gp42. A hypothesis concerning the molecular coevolution of DNA replication and recombination pathways is developed. This hypothesis is used to explain the significantly different ways in which each of the enzymatically equivalent "recA" proteins substitutes for the E. coli RecA protein and gpuvsX.
Type:
text; Dissertation-Reproduction (electronic)
Keywords:
Biology.
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Genetics; Graduate College
Degree Grantor:
University of Arizona
Advisor:
Bernstein, Harris

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleRecombination-dependent DNA replication in bacteriophage T4: An evolutionary study.en_US
dc.creatorMcCreary, Ronald Patrick.en_US
dc.contributor.authorMcCreary, Ronald Patrick.en_US
dc.date.issued1990en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractGenetic recombination is an ubiquitious process in living organisms and is one of the most elaborate activities that DNA undergoes. As such, the recombination process potentially interacts with all of the major metabolic events involving DNA. This study examined one of these interactions, specifically that occuring between recombination and DNA replication in phage T4. This interaction was investigated from both the mechanisitic and evolutionary viewpoints. In phage and bacteria, the central reactions of homologous recombination are those of homologous pairing and strand insertion. These reactions are generally catalyzed by a single enzyme. The E. coli recA gene product serves as the paradigm for this class of enzyme. Four cloned wild type "recA" genes from the bacterial species E. coli, A. caviae, B. pertussis, and the uvsX gene from phage T4, were used to test for complementation of both E. coli recA⁻ and phage T4 uvsX⁻ mutants. The B. pertussis recA⁺ gene was able to complement an E. coli recA⁻ mutant with respect to the repair of UV induced DNA damage and general recombination. In a phage Mode 2 DNA synthesis assay, all three bacterial recA genes complemented a uvsX⁻ mutant to some extent, but the B. pertussis RecA protein allowed "runaway" DNA synthesis to occur. In contrast, none of the three bacterial genes were able to increase DNA synthesis of a uvsX⁺, uvsY⁻ phage T4. In a one step growth experiment, only the B. pertussis RecA protein complemented a phage uvsX⁻ mutant. However, none of the three bacterial recA genes complemented the same phage mutant in a UV survival assay. These results imply that normal wild type Mode 2 phage DNA synthesis is an example of a specific interaction between the DNA replication and homologous recombination "machines" of phage T4. Multiplicity reactivation (MR) is a type of recombinational repair. MR experiments were performed using a gene 42(ts) mutant phage to study the effect of this member of the phage's DNA replication machinery on recombinational DNA repair. The data show that MR is dependent on the gene 42 gene product suggesting that the phage DNA replication and recombination metabolic pathways are coupled through a protein-protein interaction involving gpuvsX and gp42. A hypothesis concerning the molecular coevolution of DNA replication and recombination pathways is developed. This hypothesis is used to explain the significantly different ways in which each of the enzymatically equivalent "recA" proteins substitutes for the E. coli RecA protein and gpuvsX.en_US
dc.typetexten_US
dc.typeDissertation-Reproduction (electronic)en_US
dc.subjectBiology.en_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineGeneticsen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorBernstein, Harrisen_US
dc.contributor.committeememberWard, Oscaren_US
dc.contributor.committeememberRay, Dennisen_US
dc.contributor.committeememberIto, Junetsuen_US
dc.contributor.committeememberFreidman, Richarden_US
dc.identifier.proquest9111952en_US
dc.identifier.oclc710365991en_US
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