Attachment site selection and identity in Bxb1 serine integrase-mediated site-specific recombination.
Attachment site selection and identity in Bxb1 serine integrase-mediated site-specific recombination.
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DOI：
10.1371/journal.pgen.1003490
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发表时间：
2013-05
期刊：
影响因子：
4.5
通讯作者：
Hatfull GF
中科院分区：
文献类型：
--
作者：
Singh S;Ghosh P;Hatfull GF
Phage-encoded serine integrases mediate directionally regulated site-specific recombination between short attP and attB DNA sites without host factor requirements. These features make them attractive for genome engineering and synthetic genetics, although the basis for DNA site selection is poorly understood. Here we show that attP selection is determined through multiple proofreading steps that reject non-attP substrates, and that discrimination of attP and attB involves two critical site features: the outermost 5–6 base pairs of attP that are required for Int binding and recombination but antagonize attB function, and the “discriminators” at positions −15/+15 that determine attB identity but also antagonize attP function. Thus, although the attachment sites differ in length and sequence, only two base changes are needed to convert attP to attL, and just two more from attL to attB. The opposing effect of site identifiers ensures that site schizophrenia with dual identities does not occur. Site-specific recombinases catalyze recombination between two specific DNA sites to generate the products of recombination. The Integrase encoded by mycobacteriophage Bxb1 is a member of the serine-recombinase family and catalyzes strand exchange between attP and attB, the attachment sites for the phage and bacterial host, respectively. Although the DNA sites are relatively small (<50 bp), the reaction is highly selective for these sites and is also strongly directional. Here, we address the question of what sequences within attP are required for it to act as an attP site and identify the key sequence features that are required not just for Integrase binding but also for synapsis and post-synapsis events. We also have identified the key determinants of attP and attB identity, and although the sites are different in sequence and length, they can be interconverted with just two base changes in each of the half sites.
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影响因子：
14.9
作者：
Lewis, JA;Hatfull, GF
通讯作者：
Hatfull, GF
影响因子：
14.9
作者：
Smith, MCA;Till, R;Smith, MCM
通讯作者：
Smith, MCM
DOI：
10.1073/pnas.0711649105
发表时间：
2008-03-04
影响因子：
11.1
作者：
Ghosh, Pallavi;Bibb, Lori A.;Hatfull, Graham F.
通讯作者：
Hatfull, Graham F.
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影响因子：
14.9
作者：
Gupta, Milind;Till, Rob;Smith, Margaret C M
通讯作者：
Smith, Margaret C M
影响因子：
16
作者：
Ghosh, P;Kim, AI;Hatfull, GF
通讯作者：
Hatfull, GF