DNA Synthesis and dRPase Activities of Polymerase β Are Both Essential for Single-Nucleotide Patch Base Excision Repair in Mammalian Cell Extracts

Biochemistry ◽  
2001 ◽  
Vol 40 (3) ◽  
pp. 809-813 ◽  
Author(s):  
Andrej Ja. Podlutsky ◽  
Irina I. Dianova ◽  
Samuel H. Wilson ◽  
Vilhelm A. Bohr ◽  
Grigory L. Dianov
Keyword(s):  
Dna Synthesis ◽  
Mammalian Cell ◽  
Base Excision Repair ◽  
Excision Repair ◽  
Single Nucleotide ◽  
Cell Extracts ◽  
Base Excision

In Vitro Base Excision Repair Assay Using Mammalian Cell Extracts

1999 ◽  
pp. 301-315 ◽  
Author(s):  
Guido Frosina ◽  
Enrico Cappelli ◽  
Paola Fortini ◽  
Eugenia Dogliotti
Keyword(s):  
Mammalian Cell ◽  
Base Excision Repair ◽  
Excision Repair ◽  
Cell Extracts ◽  
Base Excision

In Vitro Base Excision Repair Assay Using Mammalian Cell Extracts

2003 ◽  
pp. 301-315 ◽  
Author(s):  
Guido Frosina ◽  
Enrico Cappelli ◽  
Paola Fortini ◽  
Eugenia Dogliotti
Keyword(s):  
Mammalian Cell ◽  
Base Excision Repair ◽  
Excision Repair ◽  
Cell Extracts ◽  
Base Excision

In Vitro Base Excision Repair Assay Using Mammalian Cell Extracts

2008 ◽  
pp. 377-396 ◽  
Author(s):  
Guido Frosina ◽  
Enrico Cappelli ◽  
Monica Ropolo ◽  
Paola Fortini ◽  
Barbara Pascucci ◽  
...  
Keyword(s):  
Mammalian Cell ◽  
Base Excision Repair ◽  
Excision Repair ◽  
Cell Extracts ◽  
Base Excision

scholarly journals Single Nucleotide Patch Base Excision Repair Is the Major Pathway for Removal of Thymine Glycol from DNA in Human Cell Extracts

2000 ◽  
Vol 275 (16) ◽  
pp. 11809-11813 ◽  
Author(s):  
Grigory L. Dianov ◽  
Tanja Thybo ◽  
Irina I. Dianova ◽  
Leonora J. Lipinski ◽  
Vilhelm A. Bohr
Keyword(s):  
Base Excision Repair ◽  
Human Cell ◽  
Excision Repair ◽  
Major Pathway ◽  
Single Nucleotide ◽  
Thymine Glycol ◽  
Cell Extracts ◽  
Base Excision

scholarly journals MSH2–MSH6 stimulates DNA polymerase η, suggesting a role for A:T mutations in antibody genes

2005 ◽  
Vol 201 (4) ◽  
pp. 637-645 ◽  
Author(s):  
Teresa M. Wilson ◽  
Alexandra Vaisman ◽  
Stella A. Martomo ◽  
Patsa Sullivan ◽  
Li Lan ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Base Excision Repair ◽  
Somatic Hypermutation ◽  
Excision Repair ◽  
Cytidine Deaminase ◽  
Abasic Site ◽  
Cell Extracts ◽  
Activation Induced Cytidine Deaminase ◽  
Base Excision

Activation-induced cytidine deaminase deaminates cytosine to uracil (dU) in DNA, which leads to mutations at C:G basepairs in immunoglobulin genes during somatic hypermutation. The mechanism that generates mutations at A:T basepairs, however, remains unclear. It appears to require the MSH2–MSH6 mismatch repair heterodimer and DNA polymerase (pol) η, as mutations of A:T are decreased in mice and humans lacking these proteins. Here, we demonstrate that these proteins interact physically and functionally. First, we show that MSH2–MSH6 binds to a U:G mismatch but not to other DNA intermediates produced during base excision repair of dUs, including an abasic site and a deoxyribose phosphate group. Second, MSH2 binds to pol η in solution, and endogenous MSH2 associates with the pol in cell extracts. Third, MSH2–MSH6 stimulates the catalytic activity of pol η in vitro. These observations suggest that the interaction between MSH2–MSH6 and DNA pol η stimulates synthesis of mutations at bases located downstream of the initial dU lesion, including A:T pairs.

scholarly journals Generation of single-nucleotide repair patches following excision of uracil residues from DNA.

1992 ◽  
Vol 12 (4) ◽  
pp. 1605-1612 ◽  
Author(s):  
G Dianov ◽  
A Price ◽  
T Lindahl
Keyword(s):  
Mammalian Cell ◽  
Excision Repair ◽  
Enzymatic Digestion ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Repair Synthesis ◽  
Single Nucleotide ◽  
Cell Extracts ◽  
Dna Repair Synthesis

The extent and location of DNA repair synthesis in a double-stranded oligonucleotide containing a single dUMP residue have been determined. Gently prepared Escherichia coli and mammalian cell extracts were employed for excision repair in vitro. The size of the resynthesized patch was estimated by restriction enzyme analysis of the repaired oligonucleotide. Following enzymatic digestion and denaturing gel electrophoresis, the extent of incorporation of radioactively labeled nucleotides in the vicinity of the lesion was determined by autoradiography. Cell extracts of E. coli and of human cell lines were shown to carry out repair mainly by replacing a single nucleotide. No significant repair replication on the 5' side of the lesion was observed. The data indicate that, after cleavage of the dUMP residue by uracil-DNA glycosylase and incision of the resultant apurinic-apyrimidinic site by an apurinic-apyrimidinic endonuclease activity, the excision step is catalyzed usually by a DNA deoxyribophosphodiesterase rather than by an exonuclease. Gap-filling and ligation complete the repair reaction. Experiments with enzyme inhibitors in mammalian cell extracts suggest that the repair replication step is catalyzed by DNA polymerase beta.

Sign in / Sign up

Export Citation Format

Share Document