Crystal structure of a thermostable type B DNA polymerase from Thermococcus gorgonarius

被引:189
作者
Hopfner, KP
Eichinger, A
Engh, RA
Laue, F
Ankenbauer, W
Huber, R
Angerer, B
机构
[1] Max Planck Inst Biochem, Abt Strukturforsch, D-82152 Martinsried, Germany
[2] Roche Diagnost, D-82372 Penzberg, Germany
关键词
x-ray structure; disulfide bonds; replication; Archaea; exonuclease;
D O I
10.1073/pnas.96.7.3600
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Most known archaeal DNA polymerases belong to the type B family, which also includes the DNA replication polymerases of eukaryotes, but maintain high fidelity at extreme conditions. We describe here the 2.5 Angstrom resolution crystal structure of a DNA polymerase from the Archaea Thermococcus gorgonarius and identify structural features of the fold and the active site that are likely responsible for its thermostable function. Comparison with the mesophilic B type DNA polymerase gp43 of the bacteriophage RB69 highlights thermophilic adaptations, which include the presence of two disulfide bonds and an enhanced electrostatic complementarity at the DNA-protein interface. In contrast to gp43, several loops in the exonuclease and thumb domains are more closely packed; this apparently blocks primer binding to the exonuclease active site. A physiological role of this "closed" conformation is unknown but may represent a polymerase mode, in contrast to an editing mode with an open exonuclease site. This archaeal 13 DNA polymerase structure provides a starting point for structure-based design of polymerases or ligands with applications in biotechnology and the development of antiviral or anticancer agents.
引用
收藏
页码:3600 / 3605
页数:6
相关论文
共 48 条
  • [21] FUNCTION AND STRUCTURE RELATIONSHIPS IN DNA-POLYMERASES
    JOYCE, CM
    STEITZ, TA
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1994, 63 : 777 - 822
  • [22] DICTIONARY OF PROTEIN SECONDARY STRUCTURE - PATTERN-RECOGNITION OF HYDROGEN-BONDED AND GEOMETRICAL FEATURES
    KABSCH, W
    SANDER, C
    [J]. BIOPOLYMERS, 1983, 22 (12) : 2577 - 2637
  • [23] DNA-POLYMERASE-III HOLOENZYME - STRUCTURE AND FUNCTION OF A CHROMOSOMAL REPLICATING MACHINE
    KELMAN, Z
    ODONNELL, M
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1995, 64 : 171 - 200
  • [24] Visualizing DNA replication in a catalytically active Bacillus DNA polymerase crystal
    Kiefer, JR
    Mao, C
    Braman, JC
    Beese, LS
    [J]. NATURE, 1998, 391 (6664) : 304 - 307
  • [25] Crystal structure of a thermostable Bacillus DNA polymerase I large fragment at 2.1 angstrom resolution
    Kiefer, JR
    Mao, C
    Hansen, CJ
    Basehore, SL
    Hogrefe, HH
    Braman, JC
    Beese, LS
    [J]. STRUCTURE, 1997, 5 (01) : 95 - 108
  • [26] CRYSTAL-STRUCTURE OF THERMUS-AQUATICUS DNA-POLYMERASE
    KIM, Y
    EOM, SH
    WANG, JM
    LEE, DS
    SUH, SW
    STEITZ, TA
    [J]. NATURE, 1995, 376 (6541) : 612 - 616
  • [27] CRYSTAL-STRUCTURE AT 3.5 ANGSTROM RESOLUTION OF HIV-1 REVERSE-TRANSCRIPTASE COMPLEXED WITH AN INHIBITOR
    KOHLSTAEDT, LA
    WANG, J
    FRIEDMAN, JM
    RICE, PA
    STEITZ, TA
    [J]. SCIENCE, 1992, 256 (5065) : 1783 - 1790
  • [28] KORNBERG A, 1992, DNA REPLICATION
  • [29] LESLIE AGW, 1994, MOSFLM USER GUIDE
  • [30] LIN TC, 1994, J BIOL CHEM, V269, P19286