SPECIFIC BINDING OF CYCLIC-AMP RECEPTOR PROTEIN TO DNA - EFFECT OF THE SEQUENCE AND OF THE INTRODUCTION OF A NICK IN THE BINDING-SITE

被引:6
作者
GIRAUDPANIS, MJ [1 ]
TOULME, F [1 ]
MAURIZOT, JC [1 ]
CULARD, F [1 ]
机构
[1] CTR BIOPHYS MOLEC,1A AVE RECH SCI,F-45071 ORLEANS 2,FRANCE
关键词
D O I
10.1080/07391102.1992.10508648
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 binding of Escherichia coli Cyclic AMP Receptor Protein (CRP) to several DNA fragments of about 45 base pairs, bearing the natural lactose or galactose sites, as well as several synthetic related sites, was investigated using fluorescence spectroscopy and gel retardation experiments. The salt dependence of the equilibrium binding constant indicates that CRP makes an identical number of ion pairs with the lac, lacL8 and gal sites although the binding constants are drastically different. However increasing the symmetry of the gal site leads to an increase of the number of ion pairs between the protein and the DNA. A single strand nick was introduced at the centre of a symmetrized gal site and this reduces the binding energy of CRP by about 0.6 Kcal. These results are discussed with respect to the bending constraints imposed on the DNA by the binding of CRP. The results are in agreement with the recently published crystal structure of the CRP complexed with DNA [Schutz, S.C., Shields, G.C. and Steitz, T.A., Science 253, 1001-1007 (1991)] showing that the 90-degrees bending of the DNA in the complex results from two kinks.
引用
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页码:295 / 309
页数:15
相关论文
共 48 条
[1]   MOLECULAR-CLONING AND NUCLEOTIDE SEQUENCING OF THE GENE FOR ESCHERICHIA-COLI CAMP RECEPTOR PROTEIN [J].
AIBA, H ;
FUJIMOTO, S ;
OZAKI, N .
NUCLEIC ACIDS RESEARCH, 1982, 10 (04) :1345-1361
[2]   A PHAGE REPRESSOR OPERATOR COMPLEX AT 7-A RESOLUTION [J].
ANDERSON, JE ;
PTASHNE, M ;
HARRISON, SC .
NATURE, 1985, 316 (6029) :596-601
[3]  
BACKMAN K, 1978, COLD SPRING HARB SYM, V43, P69
[4]   DIRECT FLUORIMETRIC MEASUREMENT OF SPECIFIC BINDING OF THE CAMP-CRP COMPLEX TO ITS SITES ON DNA FRAGMENTS OF VARIOUS LENGTH SPANNING THE CONTROL REGION OF ESCHERICHIA-COLI-K12 LACTOSE OPERON [J].
BAUDRAS, A ;
BLAZY, B ;
TAKAHASHI, M .
BIOCHIMIE, 1983, 65 (07) :437-440
[5]   SELECTION OF DNA-BINDING SITES BY REGULATORY PROTEINS .2. THE BINDING-SPECIFICITY OF CYCLIC-AMP RECEPTOR PROTEIN TO RECOGNITION SITES [J].
BERG, OG ;
VONHIPPEL, PH .
JOURNAL OF MOLECULAR BIOLOGY, 1988, 200 (04) :709-723
[6]   ORIGIN OF REPLICATION OF PBR345 PLASMID DNA [J].
BOLIVAR, F ;
BETLACH, MC ;
HEYNEKER, HL ;
SHINE, J ;
RODRIGUEZ, RL ;
BOYER, HW .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (12) :5265-5269
[7]   SEGMENT-SPECIFIC MUTAGENESIS OF THE REGULATORY REGION IN THE ESCHERICHIA-COLI GALACTOSE OPERON - ISOLATION OF MUTATIONS REDUCING THE INITIATION OF TRANSCRIPTION AND TRANSLATION [J].
BUSBY, S ;
DREYFUS, M .
GENE, 1983, 21 (1-2) :121-131
[8]   MUTATIONS IN THE ESCHERICHIA-COLI OPERON THAT DEFINE 2 PROMOTERS AND THE BINDING-SITE OF THE CYCLIC-AMP RECEPTOR PROTEIN [J].
BUSBY, S ;
AIBA, H ;
DECROMBRUGGHE, B .
JOURNAL OF MOLECULAR BIOLOGY, 1982, 154 (02) :211-227
[9]   CLONING AND SEQUENCE OF THE CRP GENE OF ESCHERICHIA-COLI K-12 [J].
COSSART, P ;
GICQUELSANZEY, B .
NUCLEIC ACIDS RESEARCH, 1982, 10 (04) :1363-1378
[10]   CYCLIC-AMP RECEPTOR PROTEIN - ROLE IN TRANSCRIPTION ACTIVATION [J].
DECROMBRUGGHE, B ;
BUSBY, S ;
BUC, H .
SCIENCE, 1984, 224 (4651) :831-838