caa a22 4500 397557345 CHVBK 20180308164806.0 cr unu---uuuuu 161202e199604 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021292 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021292 Characterization and Comparison of Synthetic Immobile and Mobile Holliday Junctions [Elektronische Daten] [Toshio Shida, Hiroshi Iwasaki, Hideo Shinagawa, Yoshimasa Kyogoku] Eight synthetic Holliday junction (HJ) oligonucleotides containing an immobile or a mobile junction were characterized by gel electrophoresis, ultraviolet absorption and circular dichroism (CD) spectroscopy. Four 24-mer deoxyribonucleotides formed stable immobile and mobile HJs in 0.1 M NaCl at 5 μM strand concentration at room temperature. However, the immobile HJ constructed from four 18-mers was less stable, and four 12-mers did not form the HJ structure under the conditions used. A comparison of the melting profiles of the HJs with those of the duplexes corresponding to the arms of four-way junctions indicated that the thermal stability of the HJ was similar to that of the individual arm and the cooperativity of the melting behavior of the HJ was relatively higher than that of the individual arm duplex. The Tms of the mobile HJs containing 4,6,8, and 10 base-pair homologous cores at junctions were essentially identical with that of the immobile HJ of the same size. There is a tendency that the HJ containing a larger homologous core region becomes more resistant to thermal denaturation. The addition of divalent metal cations, Mg2+ and Ca2+, to the solutions of the HJs raised their melting temperatures. The difference found for the CD spectra of the HJs which differ only in the arrangement of the HJ depended primarily upon the DNA sequence flanking the junction. The RuvC protein binds to the immobile and mobile HJs, regardless of the presence and the size of the homologous core at the junction. © by the Journal of Biochemistry Regular Papers nationallicence CD nationallicence Holliday junction nationallicence polyacrylamide gel electrophoresis nationallicence RuvC protein nationallicence UV nationallicence Shida Toshio Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University 3-15-1 Tokida, Ueda, Nagano 386 aut Iwasaki Hiroshi Research Institute for Microbial Diseases Osaka University 3-2 Yamadaoka, Suita, Osaka 565 aut Shinagawa Hideo Research Institute for Microbial Diseases Osaka University 3-2 Yamadaoka, Suita, Osaka 565 aut Kyogoku Yoshimasa Institute for Protein Research, Osaka University 3-2 Yamadaoka, Suita, Osaka 565 aut The Journal of Biochemistry Oxford University Press 119/4(1996-04), 653-658 0021-924X 119:4<653 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021292 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021292 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Shida Toshio Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University 3-15-1 Tokida, Ueda, Nagano 386 aut NATIONALLICENCE 700 1- Iwasaki Hiroshi Research Institute for Microbial Diseases Osaka University 3-2 Yamadaoka, Suita, Osaka 565 aut NATIONALLICENCE 700 1- Shinagawa Hideo Research Institute for Microbial Diseases Osaka University 3-2 Yamadaoka, Suita, Osaka 565 aut NATIONALLICENCE 700 1- Kyogoku Yoshimasa Institute for Protein Research, Osaka University 3-2 Yamadaoka, Suita, Osaka 565 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/4(1996-04), 653-658 0021-924X 119:4<653 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford