PMID- 23775797
OWN - NLM
STAT- MEDLINE
DCOM- 20131105
LR  - 20211203
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 41
IP  - 16
DP  - 2013 Sep
TI  - Comet-FISH with strand-specific probes reveals transcription-coupled repair of 
      8-oxoGuanine in human cells.
PG  - 7700-12
LID - 10.1093/nar/gkt524 [doi]
AB  - Oxidized bases in DNA have been implicated in cancer, aging and neurodegenerative 
      disease. We have developed an approach combining single-cell gel electrophoresis 
      (comet) with fluorescence in situ hybridization (FISH) that enables the 
      comparative quantification of low, physiologically relevant levels of DNA lesions 
      in the respective strands of defined nucleotide sequences and in the genome 
      overall. We have synthesized single-stranded probes targeting the termini of DNA 
      segments of interest using a polymerase chain reaction-based method. These probes 
      facilitate detection of damage at the single-molecule level, as the lesions are 
      converted to DNA strand breaks by lesion-specific endonucleases or glycosylases. 
      To validate our method, we have documented transcription-coupled repair of 
      cyclobutane pyrimidine dimers in the ataxia telangiectasia-mutated (ATM) gene in 
      human fibroblasts irradiated with 254 nm ultraviolet at 0.1 J/m2, a dose 
       approximately 100-fold lower than those typically used. The high specificity and sensitivity 
      of our approach revealed that 7,8-dihydro-8-oxoguanine (8-oxoG) at an incidence 
      of approximately three lesions per megabase is preferentially repaired in the 
      transcribed strand of the ATM gene. We have also demonstrated that the hOGG1, 
      XPA, CSB and UVSSA proteins, as well as actively elongating RNA polymerase II, 
      are required for this process, suggesting cross-talk between DNA repair pathways.
FAU - Guo, Jia
AU  - Guo J
AD  - Department of Biology, Stanford University, 371 Serra Mall, Stanford, CA 
      94305-5020, USA.
FAU - Hanawalt, Philip C
AU  - Hanawalt PC
FAU - Spivak, Graciela
AU  - Spivak G
LA  - eng
GR  - R01 ES018834/ES/NIEHS NIH HHS/United States
GR  - ES018834/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20130617
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Carrier Proteins)
RN  - 0 (Cell Cycle Proteins)
RN  - 0 (DNA Probes)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Poly-ADP-Ribose Binding Proteins)
RN  - 0 (Pyrimidine Dimers)
RN  - 0 (Tumor Suppressor Proteins)
RN  - 0 (UVSSA protein, human)
RN  - 5614-64-2 (8-hydroxyguanine)
RN  - 5Z93L87A1R (Guanine)
RN  - EC 2.7.11.1 (ATM protein, human)
RN  - EC 2.7.11.1 (Ataxia Telangiectasia Mutated Proteins)
RN  - EC 2.7.11.1 (Protein Serine-Threonine Kinases)
RN  - EC 2.7.7.- (RNA Polymerase II)
RN  - EC 3.6.4.- (DNA Helicases)
RN  - EC 3.6.4.12 (ERCC6 protein, human)
RN  - EC 6.5.1.- (DNA Repair Enzymes)
SB  - IM
MH  - Ataxia Telangiectasia Mutated Proteins
MH  - Carrier Proteins/physiology
MH  - Cell Cycle Proteins/biosynthesis/genetics
MH  - Cell Line
MH  - Comet Assay/*methods
MH  - DNA Helicases/physiology
MH  - DNA Probes
MH  - *DNA Repair
MH  - DNA Repair Enzymes/physiology
MH  - DNA-Binding Proteins/biosynthesis/genetics
MH  - Guanine/*analogs & derivatives/metabolism
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Poly-ADP-Ribose Binding Proteins
MH  - Protein Serine-Threonine Kinases/biosynthesis/genetics
MH  - Pyrimidine Dimers/metabolism
MH  - RNA Polymerase II/metabolism
MH  - *Transcription, Genetic
MH  - Tumor Suppressor Proteins/biosynthesis/genetics
PMC - PMC3763531
EDAT- 2013/06/19 06:00
MHDA- 2013/11/06 06:00
PMCR- 2013/06/17
CRDT- 2013/06/19 06:00
PHST- 2013/06/19 06:00 [entrez]
PHST- 2013/06/19 06:00 [pubmed]
PHST- 2013/11/06 06:00 [medline]
PHST- 2013/06/17 00:00 [pmc-release]
AID - gkt524 [pii]
AID - 10.1093/nar/gkt524 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2013 Sep;41(16):7700-12. doi: 10.1093/nar/gkt524. Epub 2013 
      Jun 17.