Lesion bypass by human DNA polymerase μ reveals a template-dependent, sequence-independent nucleotidyl transferase activity

被引:44
作者
Covo, S
Blanco, L
Livneh, Z [1 ]
机构
[1] Weizmann Inst Sci, Dept Biol Chem, IL-76100 Rehovot, Israel
[2] Univ Autonoma Madrid, CSIC, Ctr Biol Mol Severo Ochoa, E-28049 Madrid, Spain
关键词
D O I
10.1074/jbc.M310447200
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
DNA polymerase mu(pol mu), which is related to terminal deoxynucleotidyl transferase and DNA polymerase beta, is thought to be involved in non-homologous end joining and V(D) J recombination. Pol mu is induced by ionizing radiation and exhibits low fidelity. Analysis of translesion replication by purified human pol mu revealed that it bypasses a synthetic abasic site with high efficiency, using primarily a misalignment mechanism. It can also replicate across two tandem abasic sites, using the same mechanism. Pol mu extends primers whose 3'-terminal nucleotides are located opposite the abasic site. Most remarkably, this extension occurs via a mode of nucleotidyl transferase activity, which does not depend on the sequence of the template. This is not due to simple terminal nucleotidyl transferase activity, because pol mu is unable to add dNTPs to an oligo(dT)(29) primer or to a blunt end duplex oligonucleotide under standard conditions. Thus, pol mu is a dual mode DNA-synthesizing enzyme, which can act as either a classical DNA polymerase or as a non-canonical, template-dependent, but sequence-independent nucleotidyl transferase. To our knowledge, this is the first report on a DNA-synthesizing enzyme with such properties. These activities may be required for its function in non-homologous end joining in the processing of DNA ends prior to ligation.
引用
收藏
页码:859 / 865
页数:7
相关论文
共 36 条
[1]   Two novel human and mouse DNA polymerases of the polX family [J].
Aoufouchi, S ;
Flatter, E ;
Dahan, A ;
Faili, A ;
Bertocci, B ;
Storck, S ;
Delbos, F ;
Cocea, L ;
Gupta, N ;
Weill, JC ;
Reynaud, CA .
NUCLEIC ACIDS RESEARCH, 2000, 28 (18) :3684-3693
[2]   Quantitative measurement of translesion replication in human cells: Evidence for bypass of abasic sites by a replicative DNA polymerase [J].
Avkin, S ;
Adar, S ;
Blander, G ;
Livneh, Z .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (06) :3764-3769
[3]   Immunoglobulin κ light chain gene rearrangement is impaired in mice deficient for DNA polymerase mu [J].
Bertocci, B ;
De Smet, A ;
Berek, C ;
Weill, JC ;
Reynaud, CA .
IMMUNITY, 2003, 19 (02) :203-211
[4]   Cutting edge:: DNA polymerases μ and λ are dispensable for Ig gene hypermutation [J].
Bertocci, B ;
De Smet, A ;
Flatter, E ;
Dahan, A ;
Bories, JC ;
Landreau, C ;
Weill, JC ;
Reynaud, CA .
JOURNAL OF IMMUNOLOGY, 2002, 168 (08) :3702-3706
[5]   Translesion replication by DNA polymerase β is modulated by sequence context and stimulated by fork-like flap structures in DNA [J].
Daube, SS ;
Arad, G ;
Livneh, Z .
BIOCHEMISTRY, 2000, 39 (02) :397-405
[6]   Translesion replication by DNA polymerase δ depends on processivity accessory proteins and differs in specificity from DNA polymerase β [J].
Daube, SS ;
Tomer, G ;
Livneh, Z .
BIOCHEMISTRY, 2000, 39 (02) :348-355
[7]   DNA polymerase mu (Pol μ), homologous to TdT, could act as a DNA mutator in eukaryotic cells [J].
Domínguez, O ;
Ruiz, JF ;
de Lera, TL ;
García-Díaz, M ;
González, MA ;
Kirchhoff, T ;
Martínez-A, C ;
Bernad, A ;
Blanco, L .
EMBO JOURNAL, 2000, 19 (07) :1731-1742
[8]   Human DNA polymerase mu (Pol μ) exhibits an unusual replication slippage ability at AAF lesion [J].
Duvauchelle, JB ;
Blanco, L ;
Fuchs, RPP ;
Cordonnier, AM .
NUCLEIC ACIDS RESEARCH, 2002, 30 (09) :2061-2067
[9]  
Efrati E, 1997, J BIOL CHEM, V272, P2559
[10]  
FRIEDBERG EC, 1995, DNA REPAIR MUTAGENES, P19