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J Mol Biol. 2014 Jan 23;426(2):318-31. doi: 10.1016/j.jmb.2013.10.013. Epub 2013 Oct 22.

Cross-talk between diverse serine integrases.

Author information

1
Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15241, USA.
2
Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15241, USA. Electronic address: gfh@pitt.edu.

Abstract

Phage-encoded serine integrases are large serine recombinases that mediate integrative and excisive site-specific recombination of temperate phage genomes. They are well suited for use in heterologous systems and for synthetic genetic circuits as the attP and attB attachment sites are small (<50 bp), there are no host factor or DNA supercoiling requirements, and they are strongly directional, doing only excisive recombination in the presence of a recombination directionality factor. Combining different recombinases that function independently and without cross-talk to construct complex synthetic circuits is desirable, and several different serine integrases are available. However, we show here that these functions are not reliably predictable, and we describe a pair of serine integrases encoded by mycobacteriophages Bxz2 and Peaches with unusual and unpredictable specificities. The integrases share only 59% amino acid sequence identity and the attP sites have fewer than 50% shared bases, but they use the same attB site and there is non-reciprocal cross-talk between the two systems. The DNA binding specificities do not result from differences in specific DNA contacts but from the constraints imposed by the configuration of the component half-sites within each of the attachment site DNAs.

KEYWORDS:

BSA; C-terminal domain; CC; CTD; N-terminal domain; NTD; RD; ZD; attachment site selection; bovine serum albumin; coiled coil; integrase; phage integration; recombinase domain; serine recombinase; site-specific recombination; zinc-nucleated domain

PMID:
24161951
PMCID:
PMC3947336
DOI:
10.1016/j.jmb.2013.10.013
[Indexed for MEDLINE]
Free PMC Article

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