Cross-talk between Diverse Serine Integrases

JOURNAL OF MOLECULAR BIOLOGY

卷:426

Article

Singh, Shweta¹  Rockenbach, Kate¹  Dedrick, Rebekah M.¹  VanDemark, Andrew P.¹  Hatfull, Graham F.¹ 

[1] Univ Pittsburgh, Dept Biol Sci, Pittsburgh, PA 15241 USA

关键词: site-specific recombination;    serine recombinase;    integrase;    phage integration;    attachment site selection;   

DOI  :  10.1016/j.jmb.2013.10.013

来源: Elsevier

【 摘 要 】

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. (C) 2013 Elsevier Ltd. All rights reserved.

【 授权许可】

Free