A specificity switch in selected Cre recombinase variants is mediated by macromolecular plasticity and water

被引:26
作者
Baldwin, EP
Martin, SS
Abel, J
Gelato, KA
Kim, H
Schultz, PG
Santoro, SW
机构
[1] Univ Calif Davis, Sect Mol & Cellular Biol, Davis, CA 95616 USA
[2] Univ Calif Davis, Dept Chem, Davis, CA 95616 USA
[3] Davis High Sch, Yolo Cty Reg Opportun Program, Davis, CA 95616 USA
[4] Scripps Res Inst, Dept Chem, La Jolla, CA 92037 USA
[5] Scripps Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA
来源
CHEMISTRY & BIOLOGY | 2003年 / 10卷 / 11期
关键词
D O I
10.1016/j.chembiol.2003.10.015
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The basis for the altered DNA specificities of two Cre recombinase variants, obtained by mutation and selection, was revealed by their cocrystal structures. The proteins share similar substitutions but differ in their preferences for the natural LoxP substrate and an engineered substrate that is inactive with wild-type Cre, LoxM7. One variant preferentially recombines LoxM7 and contacts the substituted bases through a hydrated network of novel interlocking protein-DNA contacts. The other variant recognizes both LoxP and LoxM7 utilizing the same DNA backbone contact but different base contacts, facilitated by an unexpected DNA shift. Assisted by water, novel interaction networks can arise from few protein substitutions, suggesting how new DNA binding specificities might evolve. The contributions of macromolecular plasticity and water networks in specific DNA recognition observed here present a challenge for predictive schemes.
引用
收藏
页码:1085 / 1094
页数:10
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