Small-scale extracts for the study of nucleotide excision repair and non-homologous end joining

被引：28

作者：

Smeaton, Michael B. ^{[1
]}

Miller, Paul S. ^{[1
]}

Ketner, Gary ^{[2
]}

Hanakahi, Les A. ^{[1
]}

机构：

[1] Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Dept Biochem & Mol Biol, Baltimore, MD 21205 USA

[2] Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Dept Mol Microbiol & Immunol, Baltimore, MD 21205 USA

来源：

NUCLEIC ACIDS RESEARCH | 2007年 / 35卷 / 22期

关键词：

STRAND BREAK REPAIR; HUMAN CELL-EXTRACTS; ADENOVIRUS E4 34K; DNA; COMPLEX;

D O I：

10.1093/nar/gkm974

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 repair of DNA by nucleotide excision repair (NER) and non-homologous end joining (NHEJ) is essential for maintenance of genomic integrity and cell viability. Examination of NHEJ and NER in vitro using cell-free extracts has led to a deeper understanding of the biochemical mechanisms that underlie these processes. Current methods for production of whole-cell extracts (WCEs) to investigate NER and NHEJ start with one or more liters of culture containing 1-5x10(9) cells. Here, we describe a small-scale method for production of WCE that can be used to study NER. We also describe a rapid, small-scale method for the preparation of WCE that can be used in the study of NHEJ. These methods require less time, 20- to 1000-fold fewer cells than large-scale extracts, facilitate examination of numerous samples and are ideal for such applications as the study of host-virus interactions and analysis of mutant cell lines.

引用

页数：6

共 17 条

[1] Adenovirus E4 34k and E1b 55k oncoproteins target host DNA ligase IV for proteasomal degradation
Baker, Amy
Rohleder, Kent J.
Hanakahi, Les A.
Ketner, Gary
[J]. JOURNAL OF VIROLOGY, 2007, 81 (13) : 7034 - 7040
[2] DNA end-joining catalyzed by human cell-free extracts
Baumann, P
West, SC
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (24) : 14066 - 14070
[3] Adenovirus E4 34k and E4 11k inhibit double strand break repair and are physically associated with the cellular DNA-dependent protein kinase
Boyer, J
Rohleder, K
Ketner, G
[J]. VIROLOGY, 1999, 263 (02) : 307 - 312
[4] Role of non-homologous end joining (NHEJ) in maintaining genomic integrity
Burma, Sandeep
Chen, Benjamin P. C.
Chen, David J.
[J]. DNA REPAIR, 2006, 5 (9-10) : 1042 - 1048
[5] The Mre11 complex is required for ATM activation and the G2/M checkpoint
Carson, CT
Schwartz, RA
Stracker, TH
Lilley, CE
Lee, DV
Weitzman, MD
[J]. EMBO JOURNAL, 2003, 22 (24) : 6610 - 6620
[6] Inhibition of double-strand break non-homologous end-joining by cisplatin adducts in human cell extracts
Diggle, CP
Bentley, J
Knowles, MA
Kiltie, AE
[J]. NUCLEIC ACIDS RESEARCH, 2005, 33 (08) : 2531 - 2539
[7] DNA double strand break repair and chromosomal translocation: Lessons from animal models
Ferguson, DO
Alt, FW
[J]. ONCOGENE, 2001, 20 (40) : 5572 - 5579
[8] Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair
Hanakahi, LA
Bartlet-Jones, M
Chappell, C
Pappin, D
West, SC
[J]. CELL, 2000, 102 (06) : 721 - 729
[9] Genome maintenance mechanisms for preventing cancer
Hoeijmakers, JHJ
[J]. NATURE, 2001, 411 (6835) : 366 - 374
[10] DNA-DEPENDENT TRANSCRIPTION OF ADENOVIRUS GENES IN A SOLUBLE WHOLE-CELL EXTRACT
MANLEY, JL
FIRE, A
CANO, A
SHARP, PA
GEFTER, ML
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (07): : 3855 - 3859

← 1 2 →