Atomic force microscopy of supercoiled DNA structure on mica

被引:50
作者
Tanigawa, M [1 ]
Okada, T [1 ]
机构
[1] Joint Res Ctr Atom Technol, Ibaraki, Osaka 305, Japan
关键词
supercoiled DNA; atomic force microscopy; spermine; spermidine;
D O I
10.1016/S0003-2670(97)00709-5
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Atomic force microscopy (AFM) has been used to investigate supercoiled DNA structure. In AFM imaging, the deformation of DNA structure in the adsorption process onto the mica surface is a serious problem. To investigate the structure of supercoiled DNA in solution by AFM, the method by which DNA is fixed on the substrate without any deformation of the structure should be developed. In this paper, it is made c:lear that the structure of supercoiled DNA on mica is dependent on the method of treatment of the mica substrate. We prepared four kinds of mica surfaces; which were (1) no treatment (freshly cleaved) mica, treated with (2) spermine, (3) spermidine and (4) 3-aminopropyltriethoxysilane. The values of roughness of the mica treated with spermine and spermidine were 0.18 and 0.19 nm, respectively, while that of the mica treated with 3-aminopropyltriethoxy silane was 0.38 nm. Supercoiled DNA molecules (pUC19) were fixed in the shape of open circles on the freshly cleaved mica surface, whereas supercoiled DNA molecules on the mica treated with spermine or spermidine interwound tightly. We could, moreover, observe the clockwise handedness of the supercoiled DNA with AFM using the mica treated with spermine as substrate. The apparent length of DNA in the supercoil was shorter than that in open circular DNA. It was made clear by analysis of the differences of these measured lengths that the superhelix winding angle and the value of writhe of pUC19 were 61 degrees and 10.2, respectively. (C) 1998 Elsevier Science B.V.
引用
收藏
页码:19 / 25
页数:7
相关论文
共 22 条
[1]   DIRECT VISUALIZATION OF SUPERCOILED DNA-MOLECULES IN SOLUTION [J].
ADRIAN, M ;
TENHEGGELERBORDIER, B ;
WAHLI, W ;
STASIAK, AZ ;
STASIAK, A ;
DUBOCHET, J .
EMBO JOURNAL, 1990, 9 (13) :4551-4554
[2]  
Bates A. D., 1993, DNA topology
[3]  
Bendar J., 1994, J MOL BIOL, V235, P825
[4]   STRUCTURE OF PLECTONEMICALLY SUPERCOILED DNA [J].
BOLES, TC ;
WHITE, JH ;
COZZARELLI, NR .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 213 (04) :931-951
[5]  
BRADY GW, 1987, J MOL BIOL, V195, P184
[6]   CONFORMATIONAL VARIATION IN SUPERHELICAL DEOXYRIBONUCLEIC-ACID [J].
CAMPBELL, AM .
BIOCHEMICAL JOURNAL, 1978, 171 (01) :281-283
[7]   DNA BENDING BY CRO PROTEIN IN SPECIFIC AND NONSPECIFIC COMPLEXES - IMPLICATIONS FOR PROTEIN SITE RECOGNITION AND SPECIFICITY [J].
ERIE, DA ;
YANG, GL ;
SCHULTZ, HC ;
BUSTAMANTE, C .
SCIENCE, 1994, 266 (5190) :1562-1566
[8]   Effects of Na+ and Mg2+ on the structures of supercoiled DNAs: Comparison of simulations with experiments [J].
Gebe, JA ;
Delrow, JJ ;
Heath, PJ ;
Fujimoto, BS ;
Stewart, DW ;
Schurr, JM .
JOURNAL OF MOLECULAR BIOLOGY, 1996, 262 (02) :105-128
[9]   DNA binding to mica correlates with cationic radius: Assay by atomic force microscopy [J].
Hansma, HG ;
Laney, DE .
BIOPHYSICAL JOURNAL, 1996, 70 (04) :1933-1939
[10]   BENDING AND STRAIGHTENING OF DNA INDUCED BY THE SAME LIGAND - CHARACTERIZATION WITH THE ATOMIC-FORCE MICROSCOPE [J].
HANSMA, HG ;
BROWNE, KA ;
BEZANILLA, M ;
BRUICE, TC .
BIOCHEMISTRY, 1994, 33 (28) :8436-8441