Conferring RNA polymerase activity to a DNA polymerase: A single residue in reverse transcriptase controls substrate selection

被引:164
作者
Gao, GX [1 ]
Orlova, M [1 ]
Georgiadis, MM [1 ]
Hendrickson, WA [1 ]
Goff, SP [1 ]
机构
[1] RUTGERS STATE UNIV,WAKSMAN INST,PISCATAWAY,NJ 08855
关键词
deoxyribonucleotides; DNA synthesis;
D O I
10.1073/pnas.94.2.407
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
The traditional classification of nucleic acid polymerases as either DNA or RNA polymerases is based, in large part, on their fundamental preference for the incorporation of either deoxyribonucleotides or ribonucleotides during chain elongation. The refined structure determination of Moloney murine leukemia virus reverse transcriptase, a strict DNA polymerase, recently allowed the prediction that a single amino acid residue at the active site might be responsible for the discrimination against the 2'OH group of an incoming ribonucleotide. Mutation of this residue resulted in a variant enzyme now capable of acting as an RNA polymerase. In marked contrast to the wild-type enzyme, the K-m of the mutant enzyme for ribonucleotides was comparable to that for deoxyribonucleotides. The results are consistent with proposals of a common evolutionary origin for both classes of enzymes and support models of a common mechanism of nucleic acid synthesis underlying catalysis by all such polymerases.
引用
收藏
页码:407 / 411
页数:5
相关论文
共 31 条
  • [11] CONFORMATIONAL COUPLING IN DNA-POLYMERASE FIDELITY
    JOHNSON, KA
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1993, 62 : 685 - 713
  • [12] FUNCTION AND STRUCTURE RELATIONSHIPS IN DNA-POLYMERASES
    JOYCE, CM
    STEITZ, TA
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1994, 63 : 777 - 822
  • [13] KOHLSTAEDT LA, 1993, REVERSE TRANSCRIPTAS, P223
  • [14] Kornberg A., 1974, DNA SYNTHESIS
  • [15] KORNBERG A, 1991, DNA REPLICATION
  • [16] THE PHAGE RNA-POLYMERASES ARE RELATED TO DNA-POLYMERASES AND REVERSE TRANSCRIPTASES
    MCALLISTER, WT
    RASKIN, CA
    [J]. MOLECULAR MICROBIOLOGY, 1993, 10 (01) : 1 - 6
  • [17] STRUCTURE OF LARGE FRAGMENT OF ESCHERICHIA-COLI DNA-POLYMERASE-I COMPLEXED WITH DTMP
    OLLIS, DL
    BRICK, P
    HAMLIN, R
    XUONG, NG
    STEITZ, TA
    [J]. NATURE, 1985, 313 (6005) : 762 - 766
  • [18] INSIGHTS INTO DNA POLYMERIZATION MECHANISMS FROM STRUCTURE AND FUNCTION-ANALYSIS OF HIV-1 REVERSE-TRANSCRIPTASE
    PATEL, PH
    JACOBOMOLINA, A
    DING, JP
    TANTILLO, C
    CLARK, AD
    RAAG, R
    NANNI, RG
    HUGHES, SH
    ARNOLD, E
    [J]. BIOCHEMISTRY, 1995, 34 (16) : 5351 - 5363
  • [19] STRUCTURES OF TERNARY COMPLEXES OF RAT DNA-POLYMERASE-BETA, A DNA TEMPLATE-PRIMER, AND DDCTP
    PELLETIER, H
    SAWAYA, MR
    KUMAR, A
    WILSON, SH
    KRAUT, J
    [J]. SCIENCE, 1994, 264 (5167) : 1891 - 1903
  • [20] PRASAD VR, 1989, J BIOL CHEM, V264, P16689