The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 °C. Spies, M., Kil, Y., Masui, R., Kato, R., Kujo, C., Ohshima, T., Kuramitsu, S., & Lanzov, V. European Journal of Biochemistry, 267(4):1125-1137, 2000. cited By 22![The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 °C [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex The radA gene is an archaeal homolog of bacterial recA and eukaryotic RAD51 genes, which are critical components in homologous recombination and recombinational DNA repair. We cloned the radA gene from a hyperthermophilic archaeon, Pyrobaculum islandicum, overproduced the radA gene product in Escherichia coli and purified it to homogeneity. The purified P. islandicum RadA protein maintained its secondary structure and activities in vitro at high temperatures, up to 87 °C. It also showed high stability of 18.3 kcal·mol-1 (76.5 kJ·mol-1) at 25 °C and neutral pH. P. islandicum RadA exhibited activities typical of the family of RecA-like proteins, such as the ability to bind ssDNA, to hydrolyze ATP in a DNA-dependent manner and to catalyze DNA strand exchange. At 75 °C, all DNAs tested stimulated ATPase activity of the RadA. The protein exhibited a break in the Arrhenius plot of ATP hydrolysis at 75 °C. The cooperativity of ATP hydrolysis and ssDNA- binding ability of the protein above 75 °C were higher than at lower temperatures, and the activation energy of ATP hydrolysis was lower above this break point temperature. These results suggest that the ssDNA-dependent ATPase activity of P. islandicum RadA displays a temperature-dependent capacity to exist in two different catalytic modes, with 75 °C being the critical threshold temperature.
@ARTICLE{Spies20001125,
author={Spies, M. and Kil, Y. and Masui, R. and Kato, R. and Kujo, C. and Ohshima, T. and Kuramitsu, S. and Lanzov, V.},
title={The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 °C},
journal={European Journal of Biochemistry},
year={2000},
volume={267},
number={4},
pages={1125-1137},
doi={10.1046/j.1432-1327.2000.01108.x},
note={cited By 22},
url={https://www.scopus.com/inward/record.uri?eid=2-s2.0-0033952524&doi=10.1046%2fj.1432-1327.2000.01108.x&partnerID=40&md5=39ffd8272231ee93b715ed3af8098efd},
affiliation={Dept. of Molec. and Radiat. Biophys., Petersburg Nucl. Physics Institute, Russian Academy of Sciences, St Petersburg, Russian Federation; Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan},
abstract={The radA gene is an archaeal homolog of bacterial recA and eukaryotic RAD51 genes, which are critical components in homologous recombination and recombinational DNA repair. We cloned the radA gene from a hyperthermophilic archaeon, Pyrobaculum islandicum, overproduced the radA gene product in Escherichia coli and purified it to homogeneity. The purified P. islandicum RadA protein maintained its secondary structure and activities in vitro at high temperatures, up to 87 °C. It also showed high stability of 18.3 kcal·mol-1 (76.5 kJ·mol-1) at 25 °C and neutral pH. P. islandicum RadA exhibited activities typical of the family of RecA-like proteins, such as the ability to bind ssDNA, to hydrolyze ATP in a DNA-dependent manner and to catalyze DNA strand exchange. At 75 °C, all DNAs tested stimulated ATPase activity of the RadA. The protein exhibited a break in the Arrhenius plot of ATP hydrolysis at 75 °C. The cooperativity of ATP hydrolysis and ssDNA- binding ability of the protein above 75 °C were higher than at lower temperatures, and the activation energy of ATP hydrolysis was lower above this break point temperature. These results suggest that the ssDNA-dependent ATPase activity of P. islandicum RadA displays a temperature-dependent capacity to exist in two different catalytic modes, with 75 °C being the critical threshold temperature.},
author_keywords={Arrhenius plot; DNA-dependent ATPase; Homologous genetic recombination; Pyrobaculum islandicum; RadA},
correspondence_address1={Kuramitsu, S.; Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan; email: kuramitu@bio.sci.osaka.u.ac.jp},
issn={00142956},
coden={EJBCA},
pubmed_id={10672022},
language={English},
abbrev_source_title={Eur. J. Biochem.},
document_type={Article},
source={Scopus},
}
Downloads: 0
{"_id":"k6nkAbSGyq8ArGQm5","bibbaseid":"spies-kil-masui-kato-kujo-ohshima-kuramitsu-lanzov-theradaproteinfromahyperthermophilicarchaeonpyrobaculumislandicumisadnadependentatpasethatexhibitstwodisparatecatalyticmodeswithatransitiontemperatureat75c-2000","author_short":["Spies, M.","Kil, Y.","Masui, R.","Kato, R.","Kujo, C.","Ohshima, T.","Kuramitsu, S.","Lanzov, V."],"bibdata":{"bibtype":"article","type":"article","author":[{"propositions":[],"lastnames":["Spies"],"firstnames":["M."],"suffixes":[]},{"propositions":[],"lastnames":["Kil"],"firstnames":["Y."],"suffixes":[]},{"propositions":[],"lastnames":["Masui"],"firstnames":["R."],"suffixes":[]},{"propositions":[],"lastnames":["Kato"],"firstnames":["R."],"suffixes":[]},{"propositions":[],"lastnames":["Kujo"],"firstnames":["C."],"suffixes":[]},{"propositions":[],"lastnames":["Ohshima"],"firstnames":["T."],"suffixes":[]},{"propositions":[],"lastnames":["Kuramitsu"],"firstnames":["S."],"suffixes":[]},{"propositions":[],"lastnames":["Lanzov"],"firstnames":["V."],"suffixes":[]}],"title":"The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 °C","journal":"European Journal of Biochemistry","year":"2000","volume":"267","number":"4","pages":"1125-1137","doi":"10.1046/j.1432-1327.2000.01108.x","note":"cited By 22","url":"https://www.scopus.com/inward/record.uri?eid=2-s2.0-0033952524&doi=10.1046%2fj.1432-1327.2000.01108.x&partnerID=40&md5=39ffd8272231ee93b715ed3af8098efd","affiliation":"Dept. of Molec. and Radiat. Biophys., Petersburg Nucl. Physics Institute, Russian Academy of Sciences, St Petersburg, Russian Federation; Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan","abstract":"The radA gene is an archaeal homolog of bacterial recA and eukaryotic RAD51 genes, which are critical components in homologous recombination and recombinational DNA repair. We cloned the radA gene from a hyperthermophilic archaeon, Pyrobaculum islandicum, overproduced the radA gene product in Escherichia coli and purified it to homogeneity. The purified P. islandicum RadA protein maintained its secondary structure and activities in vitro at high temperatures, up to 87 °C. It also showed high stability of 18.3 kcal·mol-1 (76.5 kJ·mol-1) at 25 °C and neutral pH. P. islandicum RadA exhibited activities typical of the family of RecA-like proteins, such as the ability to bind ssDNA, to hydrolyze ATP in a DNA-dependent manner and to catalyze DNA strand exchange. At 75 °C, all DNAs tested stimulated ATPase activity of the RadA. The protein exhibited a break in the Arrhenius plot of ATP hydrolysis at 75 °C. The cooperativity of ATP hydrolysis and ssDNA- binding ability of the protein above 75 °C were higher than at lower temperatures, and the activation energy of ATP hydrolysis was lower above this break point temperature. These results suggest that the ssDNA-dependent ATPase activity of P. islandicum RadA displays a temperature-dependent capacity to exist in two different catalytic modes, with 75 °C being the critical threshold temperature.","author_keywords":"Arrhenius plot; DNA-dependent ATPase; Homologous genetic recombination; Pyrobaculum islandicum; RadA","correspondence_address1":"Kuramitsu, S.; Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan; email: kuramitu@bio.sci.osaka.u.ac.jp","issn":"00142956","coden":"EJBCA","pubmed_id":"10672022","language":"English","abbrev_source_title":"Eur. J. Biochem.","document_type":"Article","source":"Scopus","bibtex":"@ARTICLE{Spies20001125,\r\nauthor={Spies, M. and Kil, Y. and Masui, R. and Kato, R. and Kujo, C. and Ohshima, T. and Kuramitsu, S. and Lanzov, V.},\r\ntitle={The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 °C},\r\njournal={European Journal of Biochemistry},\r\nyear={2000},\r\nvolume={267},\r\nnumber={4},\r\npages={1125-1137},\r\ndoi={10.1046/j.1432-1327.2000.01108.x},\r\nnote={cited By 22},\r\nurl={https://www.scopus.com/inward/record.uri?eid=2-s2.0-0033952524&doi=10.1046%2fj.1432-1327.2000.01108.x&partnerID=40&md5=39ffd8272231ee93b715ed3af8098efd},\r\naffiliation={Dept. of Molec. and Radiat. Biophys., Petersburg Nucl. Physics Institute, Russian Academy of Sciences, St Petersburg, Russian Federation; Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan},\r\nabstract={The radA gene is an archaeal homolog of bacterial recA and eukaryotic RAD51 genes, which are critical components in homologous recombination and recombinational DNA repair. We cloned the radA gene from a hyperthermophilic archaeon, Pyrobaculum islandicum, overproduced the radA gene product in Escherichia coli and purified it to homogeneity. The purified P. islandicum RadA protein maintained its secondary structure and activities in vitro at high temperatures, up to 87 °C. It also showed high stability of 18.3 kcal·mol-1 (76.5 kJ·mol-1) at 25 °C and neutral pH. P. islandicum RadA exhibited activities typical of the family of RecA-like proteins, such as the ability to bind ssDNA, to hydrolyze ATP in a DNA-dependent manner and to catalyze DNA strand exchange. At 75 °C, all DNAs tested stimulated ATPase activity of the RadA. The protein exhibited a break in the Arrhenius plot of ATP hydrolysis at 75 °C. The cooperativity of ATP hydrolysis and ssDNA- binding ability of the protein above 75 °C were higher than at lower temperatures, and the activation energy of ATP hydrolysis was lower above this break point temperature. These results suggest that the ssDNA-dependent ATPase activity of P. islandicum RadA displays a temperature-dependent capacity to exist in two different catalytic modes, with 75 °C being the critical threshold temperature.},\r\nauthor_keywords={Arrhenius plot; DNA-dependent ATPase; Homologous genetic recombination; Pyrobaculum islandicum; RadA},\r\ncorrespondence_address1={Kuramitsu, S.; Department of Biology, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka 560-0043, Japan; email: kuramitu@bio.sci.osaka.u.ac.jp},\r\nissn={00142956},\r\ncoden={EJBCA},\r\npubmed_id={10672022},\r\nlanguage={English},\r\nabbrev_source_title={Eur. J. Biochem.},\r\ndocument_type={Article},\r\nsource={Scopus},\r\n}\r\n\r\n","author_short":["Spies, M.","Kil, Y.","Masui, R.","Kato, R.","Kujo, C.","Ohshima, T.","Kuramitsu, S.","Lanzov, V."],"key":"Spies20001125","id":"Spies20001125","bibbaseid":"spies-kil-masui-kato-kujo-ohshima-kuramitsu-lanzov-theradaproteinfromahyperthermophilicarchaeonpyrobaculumislandicumisadnadependentatpasethatexhibitstwodisparatecatalyticmodeswithatransitiontemperatureat75c-2000","role":"author","urls":{"Paper":"https://www.scopus.com/inward/record.uri?eid=2-s2.0-0033952524&doi=10.1046%2fj.1432-1327.2000.01108.x&partnerID=40&md5=39ffd8272231ee93b715ed3af8098efd"},"metadata":{"authorlinks":{}}},"bibtype":"article","biburl":"https://bio.pnpi.nrcki.ru/wp-content/uploads/2019/12/lmkb_2019_10.txt","dataSources":["rgjRz8LhiuL9Prc4S"],"keywords":[],"search_terms":["rada","protein","hyperthermophilic","archaeon","pyrobaculum","islandicum","dna","dependent","atpase","exhibits","two","disparate","catalytic","modes","transition","temperature","spies","kil","masui","kato","kujo","ohshima","kuramitsu","lanzov"],"title":"The RadA protein from a hyperthermophilic archaeon Pyrobaculum islandicum is a DNA-dependent ATPase that exhibits two disparate catalytic modes, with a transition temperature at 75 °C","year":2000}