Dimerization of bacteriophage P2 integrase is not required for binding to its DNA target but for its biological activity

Frumerie, Clara; Eriksson, Jesper M.; Dugast, Marc; Haggård‐Ljungquist, Elisabeth

doi:10.1016/j.gene.2004.09.026

Cited by 5 publications

(2 citation statements)

References 30 publications

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“…The glutamic acid at position 197 has previously been studied, and the E197A and E197K substitutions were proposed to cause a dimerization deficient P2 Int [26]. However, our crystal structure suggests this amino acid to also be important for stabilization of arginine residues of the DNA binding motif, as salt bridges are formed to both Arg222 and Arg194, and the distance to the nearest atom of the second subunit exceeds 12 Å.…”

Section: Resultsmentioning

confidence: 72%

“…The character of this residue is also conserved throughout integrases, with it presented as either an aspartic acid or glutamic acid, both capable of stabilizing the positively charged arginines in the near proximity. The E197A variant of P2 Int is still capable of binding attP with the same capacity as wild type P2 Int and is capable of forming phages in a complementation assay [26]. The loss of activity, both in DNA binding and the complementation assay, caused by the E197K mutation could be attributed to the charge repulsion between the introduced lysine and Arg222 and Arg194.…”

Section: Resultsmentioning

confidence: 97%

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Crystal structure of the bacteriophage P2 integrase catalytic domain

et al. 2015

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a b s t r a c tBacteriophage P2 is a temperate phage capable of integrating its DNA into the host genome by sitespecific recombination upon lysogenization. Integration and excision of the phage genome requires P2 integrase, which performs recognition, cleavage and joining of DNA during these processes. This work presents the high-resolution crystal structure of the catalytic domain of P2 integrase, and analysis of the structure-function relationship of several previously identified non-functional P2 integrase mutants. The DNA binding area is characterized by a large positively charged patch, harboring key residues. The structure reveals potential for large dimer flexibility, likely essential for rearrangement of DNA strands upon integration and excision of the phage DNA.

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Section: Resultsmentioning

confidence: 72%

Section: Resultsmentioning

confidence: 97%

Crystal structure of the bacteriophage P2 integrase catalytic domain

et al. 2015

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Bacteriophage Protein–Protein Interactions

Häuser¹,

Blasche²,

Dokland³

et al. 2012

Advances in Virus Research

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A comparative analysis of the bifunctional Cox proteins of two heteroimmune P2-like phages with different host integration sites

et al. 2009

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The Cox protein of the coliphage P2 is multifunctional; it acts as a transcriptional repressor of the Pc promoter, as a transcriptional activator of the P(LL) promoter of satellite phage P4, and as a directionality factor for site-specific recombination. The Cox proteins constitute a unique group of directionality factors since they couple the developmental switch with the integration or excision of the phage genome. In this work, the DNA binding characteristics of the Cox protein of WPhi, a P2-related phage, are compared with those of P2 Cox. P2 Cox has been shown to recognize a 9 bp sequence, repeated at least 6 times in different targets. In contrast to P2 Cox, WPhi Cox binds with a strong affinity to the early control region that contains an imperfect direct repeat of 12 nucleotides. The removal of one of the repeats has drastic effects on the capacity of WPhi to bind to the Pe-Pc region. Again in contrast to P2 Cox, WPhi Cox has a lower affinity to attP compared to the Pe-Pc region, and a repeat of 9 bp can be found that has 5 bp in common with the repeat in the Pe-Pc region. WPhi Cox, however, is essential for excisive recombination in vitro. WPhi Cox, like P2 Cox, binds cooperatively with integrase to attP. Both Cox proteins induce a strong bend in their DNA targets upon binding.

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Dimerization of bacteriophage P2 integrase is not required for binding to its DNA target but for its biological activity

Cited by 5 publications

References 30 publications

Crystal structure of the bacteriophage P2 integrase catalytic domain

Crystal structure of the bacteriophage P2 integrase catalytic domain

Bacteriophage Protein–Protein Interactions

A comparative analysis of the bifunctional Cox proteins of two heteroimmune P2-like phages with different host integration sites

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