ATYPICAL DNA-BINDING PROPERTIES OF CLASS-IIS RESTRICTION ENDONUCLEASES - EVIDENCE FOR RECOGNITION OF THE COGNATE SEQUENCE BY A FOKI MONOMER

被引：35

作者：

SKOWRON, P ^{[1
]}

KACZOROWSKI, T ^{[1
]}

TUCHOLSKI, J ^{[1
]}

PODHAJSKA, AJ ^{[1
]}

机构：

[1] UNIV GDANSK,DEPT MICROBIOL,UL KLADKI 24,PL-80822 GDANSK,POLAND

来源：

GENE | 1993年 / 125卷 / 01期

关键词：

PROTEIN DNA INTERACTION; GEL-MOBILITY-SHIFT ASSAY; ENZYME; RESTRICTION-MODIFICATION SYSTEM;

D O I：

10.1016/0378-1119(93)90738-O

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The DNA-binding properties of the FokI restriction endonuclease were studied using the gel-mobility-shift assay. Specific recognition of the cognate sequence and cleavage of DNA are distinguishable functions and can be separated. FokI binds to its recognition site predominantly as a monomer. At high concentrations, FokI exhibits a cooperative recognition sequence-dependent aggregation. In 20 mM KCl/10 mM Tris.HCl buffer, the binding constant of FokI to its cognate site is equal 6.0-7.9 x 10(8)/mol and is lower than the values for most gene-regulatory proteins. FokI binding is 600-1500 times weaker to non-cognate double-stranded DNA than to the GGATG site, and 30 000 times weaker to single-stranded DNA or tRNA. The method of Bading [Nucleic Acids Res. 16 (1988) 5241-5248], used for determining the stoichiometry of protein bound to DNA by gel-mobility-shift assay, is extended.

引用

页码：1 / 10

页数：10

共 30 条

[1]

ADAMS RLP, 1985, MOL BIOL DNA METHYLA, P44

[2] DETERMINATION OF THE MOLECULAR-WEIGHT OF DNA-BOUND PROTEIN(S) RESPONSIBLE FOR GEL-ELECTROPHORETIC MOBILITY SHIFT OF LINEAR DNA FRAGMENTS EXEMPLIFIED WITH PURIFIED VIRAL MYB PROTEIN [J].

BADING, H .

NUCLEIC ACIDS RESEARCH, 1988, 16 (12) :5241-5248

[3]

BRENNAN RG, 1989, J BIOL CHEM, V264, P1903

[4]

Bron S, 1980, Methods Enzymol, V65, P112

[5] GEL RETARDATION AT LOW PH RESOLVES TRP REPRESSOR-DNA COMPLEXES FOR QUANTITATIVE STUDY [J].

CAREY, J .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (04) :975-979

[6] EQUILIBRIUM STUDIES OF THE CYCLIC-AMP RECEPTOR PROTEIN-DNA INTERACTION [J].

FRIED, MG ;

CROTHERS, DM .

JOURNAL OF MOLECULAR BIOLOGY, 1984, 172 (03) :241-262

[7] GENETIC-ENGINEERING OF ECORI MUTANTS WITH ALTERED AMINO-ACID RESIDUES IN THE DNA-BINDING SITE - PHYSICOCHEMICAL INVESTIGATIONS GIVE EVIDENCE FOR AN ALTERED MONOMER DIMER EQUILIBRIUM FOR THE GLN144LYS145 AND GLN144LYS145LYS200 MUTANTS [J].

GEIGER, R ;

RUTER, T ;

ALVES, J ;

FLIESS, A ;

WOLFES, H ;

PINGOUD, V ;

URBANKE, C ;

MAASS, G ;

PINGOUD, A ;

DUSTERHOFT, A ;

KROGER, M .

BIOCHEMISTRY, 1989, 28 (06) :2667-2677

[8]

HALVORSEN YDC, 1990, J BIOL CHEM, V265, P13283

[9]

HARLOW E, 1988, ANTIBODIES LABORATOR

[10] LAMBDA-REPRESSOR AND CRO - COMPONENTS OF AN EFFICIENT MOLECULAR SWITCH [J].

JOHNSON, AD ;

POTEETE, AR ;

LAUER, G ;

SAUER, RT ;

ACKERS, GK ;

PTASHNE, M .

NATURE, 1981, 294 (5838) :217-223

← 1 2 3 →