Amino acid residues in Rag1 crucial for DNA hairpin formation

被引:28
作者
Lu, Catherine P.
Sandoval, Hector
Brandt, Vicky L.
Rice, Phoebe A.
Roth, David B. [1 ]
机构
[1] NYU, Sch Med, Program Mol Pathogenesis, Skirball Inst Biomol Med, New York, NY 10016 USA
[2] NYU, Sch Med, Dept Pathol, New York, NY 10016 USA
[3] Baylor Coll Med, Dept Immunol, Houston, TX 77030 USA
[4] Univ Chicago, Dept Biochem & Mol Biol, Chicago, IL 60637 USA
关键词
D O I
10.1038/nsmb1154
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 Rag proteins carry out V(D)J recombination through a process mechanistically similar to cut-and-paste transposition. Specifically, Rag complexes form DNA hairpins through direct transesterification, using a catalytic Asp-Asp-Glu (DDE) triad in Rag1. How is sufficient DNA distortion introduced to allow hairpin formation? We hypothesized that, like certain transposases, the Rag proteins might use aromatic amino acid residues to stabilize a flipped-out base. Through in vivo and in vitro experiments and structural predictions, we identified residues in Rag1 crucial for hairpin formation. One of these, a conserved tryptophan (Trp893), probably participates in base-stacking interactions near the cleavage site, as do Trp298, Trp265 and Trp319 in the Tn5, Tn10 and Hermes transposases, respectively. Other residues surrounding the catalytic glutamate (YK (E) under bar FRK) may share functional similarities with the YREK motif in IS4 family transposases.
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收藏
页码:1010 / 1015
页数:6
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