Recognition and processing of the origin of transfer DNA by conjugative relaxase TrwC

被引:121
作者
Guasch, A
Lucas, M
Moncalián, G
Cabezas, M
Pérez-Luque, R
Gomis-Rüth, FX
de la Cruz, F
Coll, M
机构
[1] CSIC, Inst Biol Mol Barcelona, ES-08034 Barcelona, Spain
[2] Univ Cantabria, CSIC, Lab CIB, Dept Biol Mol, Santander 39011, Spain
关键词
D O I
10.1038/nsb1017
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
Relaxases are DNA strand transferases that catalyze the initial and final stages of DNA processing during conjugative cell-to-cell DNA transfer. Upon binding to the origin of transfer (oriT) DNA, relaxase TrwC melts the double helix. The three-dimensional structure of the relaxase domain of TrwC in complex with its cognate DNA at oriT shows a fold built on a two-layer alpha/beta sandwich, with a deep narrow cleft that houses the active site. The DNA includes one arm of an extruded cruciform, an essential feature for specific recognition. This arm is firmly embraced by the protein through a beta-ribbon positioned in the DNA major groove and a loop occupying the minor groove. It is followed by a single-stranded DNA segment that enters the active site, after a sharp U-turn forming a hydrophobic cage that traps the N-terminal methionine. Structural analysis combined with site-directed mutagenesis defines the architecture of the active site.
引用
收藏
页码:1002 / 1010
页数:9
相关论文
共 48 条
[1]   THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY [J].
BAILEY, S .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 :760-763
[2]   Crystallography & NMR system:: A new software suite for macromolecular structure determination [J].
Brunger, AT ;
Adams, PD ;
Clore, GM ;
DeLano, WL ;
Gros, P ;
Grosse-Kunstleve, RW ;
Jiang, JS ;
Kuszewski, J ;
Nilges, M ;
Pannu, NS ;
Read, RJ ;
Rice, LM ;
Simonson, T ;
Warren, GL .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1998, 54 :905-921
[3]  
BUDISA N, 1995, EUR J BIOCHEM, V230, P788
[4]   The structure of a replication initiator unites diverse aspects of nucleic acid metabolism [J].
Campos-Olivas, R ;
Louis, JM ;
Clérot, D ;
Gronenborn, B ;
Gronenborn, AM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (16) :10310-10315
[5]   DNA topoisomerases: Structure, function, and mechanism [J].
Champoux, JJ .
ANNUAL REVIEW OF BIOCHEMISTRY, 2001, 70 :369-413
[6]   Crystal structure of a complex of a type IA DNA topoisomerase with a single-stranded DNA molecule [J].
Changela, A ;
DiGate, RJ ;
Mondragón, A .
NATURE, 2001, 411 (6841) :1077-1081
[7]   Identification of active site residues in Escherichia coli DNA topoisomerase I [J].
Chen, SJ ;
Wang, JC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (11) :6050-6056
[9]   Structure of the DNA-bound T-box domain of human TBX3, a transcription factor responsible for ulnar-mammary syndrome [J].
Coll, M ;
Seidman, JG ;
Müller, CW .
STRUCTURE, 2002, 10 (03) :343-356
[10]   Phase combination and cross validation in iterated density-modification calculations [J].
Cowtan, KD ;
Main, P .
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1996, 52 :43-48