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The Bxb1 gp47 recombination directionality factor is required not only for prophage excision, but also for phage DNA replication

Savinov, A and Pan, J and Ghosh, P and Hatfull, GF (2012) The Bxb1 gp47 recombination directionality factor is required not only for prophage excision, but also for phage DNA replication. Gene, 495 (1). 42 - 48. ISSN 0378-1119

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Abstract

Mycobacteriophage Bxb1 encodes a serine-integrase that catalyzes both integrative and excisive site-specific recombination. However, excision requires a second phage-encoded protein, gp47, which serves as a recombination directionality factor (RDF). The viability of a Bxb1 mutant containing an S153A substitution in gp47 that eliminates the RDF activity of Bxb1 gp47 shows that excision is not required for Bxb1 lytic growth. However, the inability to construct a δ47 deletion mutant of Bxb1 suggests that gp47 provides a second function that is required for lytic growth, although the possibility of an essential cis-acting site cannot be excluded. Characterization of a mutant prophage of mycobacteriophage L5 in which gene 54 - a homologue of Bxb1 gene 47 - is deleted shows that it also is defective in induced lytic growth, and exhibits a strong defect in DNA replication. Bxb1 gp47 and its relatives are also unusual in containing conserved motifs associated with a phosphoesterase function, although we have not been able to show robust phosphoesterase activity of the proteins, and amino acid substitutions with the conserved motifs do not interfere with RDF activity. We therefore propose that Bxb1 gp47 and its relatives provide an important function in phage DNA replication that has been co-opted by the integration machinery of the serine-integrases to control the directionality of recombination. © 2011 Elsevier B.V.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Savinov, A
Pan, J
Ghosh, P
Hatfull, GFgfh@pitt.eduGFH
Date: 1 March 2012
Date Type: Publication
Journal or Publication Title: Gene
Volume: 495
Number: 1
Page Range: 42 - 48
DOI or Unique Handle: 10.1016/j.gene.2011.12.003
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0378-1119
MeSH Headings: Amino Acid Sequence; Base Sequence; DNA Replication--genetics; Integrases--genetics; Integrases--physiology; Molecular Sequence Data; Mycobacteriophages--enzymology; Mycobacteriophages--genetics; Recombination, Genetic; Sequence Deletion; Virus Activation--genetics; Virus Replication--genetics
Other ID: NLM NIHMS346955 [Available on 03/01/13], NLM PMC3273658 [Available on 03/01/13]
PubMed Central ID: PMC3273658
PubMed ID: 22227494
Date Deposited: 14 Jan 2013 15:45
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16872

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