Dpo4 is hindered in extending a G•T mismatch by a reverse wobble

被引:64
作者
Trincao, J
Johnson, RE
Wolfle, WT
Escalante, CR
Prakash, S
Prakash, L
Aggarwal, AK
机构
[1] CUNY Mt Sinai Sch Med, Dept Physiol & Biophys, Struct Biol Program, New York, NY 10029 USA
[2] Univ Texas, Med Branch, Sealy Ctr Mol Sci, Galveston, TX 77555 USA
关键词
D O I
10.1038/nsmb755
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 ability or inability of a DNA polymerase to extend a mispair directly affects the establishment of genomic mutations. We report here kinetic analyses of the ability of Dpo4, a Y-family polymerase from Sulfolobus solfataricus, to extend from all mispairs opposite a template G or T. Dpo4 is equally inefficient at extending these mispairs, which include, surprisingly, a G.T mispair expected to conform closely to Watson-Crick geometry. To elucidate the basis of this, we solved the structure of Dpo4 bound to G.T-mispaired primer template in the presence of an incoming nucleotide. As a control, we also determined the structure of Dpo4 bound to a matched A-T base pair at the primer terminus. The structures offer a basis for the low efficiency of Dpo4 in extending a G.T mispair: a reverse wobble that deflects the primer 3'-OH away from the incoming nucleotide.
引用
收藏
页码:457 / 462
页数:6
相关论文
共 32 条
[1]  
AGGARWAL A K, 1990, Methods (Orlando), V1, P83, DOI 10.1016/S1046-2023(05)80150-1
[2]   The complete atomic structure of the large ribosomal subunit at 2.4 Å resolution [J].
Ban, N ;
Nissen, P ;
Hansen, J ;
Moore, PB ;
Steitz, TA .
SCIENCE, 2000, 289 (5481) :905-920
[3]  
Brunger AT, 1998, ACTA CRYSTALLOGR D, V54, P905, DOI 10.1107/s0907444998003254
[4]  
Bullock TL, 2000, NAT STRUCT BIOL, V7, P497
[5]   An open and closed case for all polymerases [J].
Doublié, S ;
Sawaya, MR ;
Ellenberger, T .
STRUCTURE, 1999, 7 (02) :R31-R35
[6]   Error-prone repair DNA polymerases in prokaryotes and eukaryotes [J].
Goodman, MF .
ANNUAL REVIEW OF BIOCHEMISTRY, 2002, 71 :17-50
[7]   Role of human DNA polymerase κ as an extender in translesion synthesis [J].
Haracska, L ;
Prakash, L ;
Prakash, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (25) :16000-16005
[8]   WOBBLE DG.DT PAIRING IN RIGHT-HANDED DNA - SOLUTION CONFORMATION OF THE D(C-G-T-G-A-A-T-T-C-G-C-G) DUPLEX DEDUCED FROM DISTANCE GEOMETRY ANALYSIS OF NUCLEAR OVERHAUSER EFFECT SPECTRA [J].
HARE, D ;
SHAPIRO, L ;
PATEL, DJ .
BIOCHEMISTRY, 1986, 25 (23) :7445-7456
[9]   EXTENSION OF BASE MISPAIRS BY TAQ DNA-POLYMERASE - IMPLICATIONS FOR SINGLE NUCLEOTIDE DISCRIMINATION IN PCR [J].
HUANG, MM ;
ARNHEIM, N ;
GOODMAN, MF .
NUCLEIC ACIDS RESEARCH, 1992, 20 (17) :4567-4573
[10]   REFINED CRYSTAL-STRUCTURE OF AN OCTANUCLEOTIDE DUPLEX WITH G.T MISMATCHED BASE-PAIRS [J].
HUNTER, WN ;
KNEALE, G ;
BROWN, T ;
RABINOVICH, D ;
KENNARD, O .
JOURNAL OF MOLECULAR BIOLOGY, 1986, 190 (04) :605-618