Molecular analyses of DNA helicases involved in the replicational stress response

被引:5
作者
Wu, Yuliang [1 ]
Sommers, Joshua A. [1 ]
Suhasini, Avvaru N. [1 ]
Aggarwal, Monika [1 ]
Brosh, Robert M. [1 ]
机构
[1] NIA, Lab Mol Gerontol, NIH, NIH Biomed Res Ctr, Baltimore, MD 21224 USA
关键词
Helicase; DNA repair; Replication; Fluorescence; ATPase; Quadruplex; Triplex; Streptavidin; FANCJ; Werner syndrome; TRIPLE-HELIX; RECQ HELICASES; FANCJ HELICASE; BIOCHEMICAL-CHARACTERIZATION; GENOMIC STABILITY; MOTOR ATPASE; IN-VIVO; MECHANISMS; OLIGONUCLEOTIDES; PROTEIN;
D O I
10.1016/j.ymeth.2010.02.021
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The importance of helicases in nucleic acid metabolism and human disease has raised the bar for understanding how these unique enzymes function to perform their biological roles at the molecular level. Here we will describe experimental procedures and strategies to investigate the functions of helicases. These functional assays have been used to study DNA helicases important for the maintenance of genomic stability and genetically linked to age-related diseases and cancer. We will focus on the description of fluorometric helicase assays, protein displacement assays, and methods to characterize helicase activity on alternate DNA structures (triplex and quadruplex) used by our laboratory. The procedures to study these helicase functions are described in step-by-step detail to enable researchers interested in nucleic acid metabolism and related fields to apply these techniques to their own research questions. Published by Elsevier Inc.
引用
收藏
页码:303 / 312
页数:10
相关论文
共 42 条
[1]   Triplex DNA in the nucleus: Direct binding of triplex-specific antibodies and their effect on transcription, replication and cell growth [J].
Agazie, YM ;
Burkholder, GD ;
Lee, JS .
BIOCHEMICAL JOURNAL, 1996, 316 :461-466
[2]   Hitting the Bull's Eye: Novel Directed Cancer Therapy Through Helicase-Targeted Synthetic Lethality [J].
Aggarwal, Monika ;
Brosh, Robert M., Jr. .
JOURNAL OF CELLULAR BIOCHEMISTRY, 2009, 106 (05) :758-763
[3]   SINGLE-TURNOVER KINETICS OF HELICASE-CATALYZED DNA UNWINDING MONITORED CONTINUOUSLY BY FLUORESCENCE ENERGY-TRANSFER [J].
BJORNSON, KP ;
AMARATUNGA, M ;
MOORE, KJM ;
LOHMAN, TM .
BIOCHEMISTRY, 1994, 33 (47) :14306-14316
[4]   Biochemical characterization of the DNA substrate specificity of Werner syndrome helicase [J].
Brosh, RM ;
Waheed, J ;
Sommers, JA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (26) :23236-23245
[5]   Biochemical characterization of the WRN-FEN-1 functional interaction [J].
Brosh, RM ;
Driscoll, HC ;
Dianov, GL ;
Sommers, JA .
BIOCHEMISTRY, 2002, 41 (40) :12204-12216
[6]   Unwinding of a DNA triple helix by the Werner and Bloom syndrome helicases [J].
Brosh, RM ;
Majumdar, A ;
Desai, S ;
Hickson, ID ;
Bohr, VA ;
Seidman, MM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (05) :3024-3030
[7]   Human premature aging, DNA repair and RecQ helicases [J].
Brosh, Robert M., Jr. ;
Bohr, Vilhelm A. .
NUCLEIC ACIDS RESEARCH, 2007, 35 (22) :7527-7544
[8]   Novel pro- and anti-recombination activities of the Bloom's syndrome helicase [J].
Bugreev, Dmitry V. ;
Yu, Xiong ;
Egelman, Edward H. ;
Mazin, Alexander V. .
GENES & DEVELOPMENT, 2007, 21 (23) :3085-3094
[9]   E-coli Rep oligomers are required to initiate DNA unwinding in vitro [J].
Cheng, W ;
Hsieh, J ;
Brendza, KM ;
Lohman, TM .
JOURNAL OF MOLECULAR BIOLOGY, 2001, 310 (02) :327-350
[10]   Biochemical and kinetic characterization of the DNA helicase and exonuclease activities of Werner syndrome protein [J].
Choudhary, S ;
Sommers, JA ;
Brosh, RM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (33) :34603-34613