Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus

Abstract All cellular organisms encode type IA topoisomerases which catalyze DNA topological changes essential for DNA transactions. However, the kinetics of the reaction catalyzed by these enzymes remains poorly characterized. Here we measured the rapid kinetics of template binding, cleavage and re...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Junhua Zhang, Bailong Pan, Zhimeng Li, Xin Sheng Zhao, Li Huang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/801302d679f649ae88b2e36adeb5d4dc
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:801302d679f649ae88b2e36adeb5d4dc
record_format	dspace
spelling	oai:doaj.org-article:801302d679f649ae88b2e36adeb5d4dc2021-12-02T11:52:26ZKinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus10.1038/s41598-017-05837-52045-2322https://doaj.org/article/801302d679f649ae88b2e36adeb5d4dc2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-05837-5https://doaj.org/toc/2045-2322Abstract All cellular organisms encode type IA topoisomerases which catalyze DNA topological changes essential for DNA transactions. However, the kinetics of the reaction catalyzed by these enzymes remains poorly characterized. Here we measured the rapid kinetics of template binding, cleavage and religation by Sso topo III, a type IA topoisomerase from the hyperthermophilic archaeon Sulfolobus solfataricus, by using a novel FRET/PIFE-based method in a stopped-flow spectrometer. We show that Sso topo III bound the template rapidly, and the rate of binding was 2–3 orders of magnitudes higher than that of template cleavage at 25 °C. The rate of template cleavage was favored over that of template religation by the enzyme, and was more so at lower temperatures (25–55 °C). Significant template cleavage [(2.23 ± 0.11) × 10−3 s−1] was observed while little religation was detectable at 25 °C. This is consistent with the presence of a higher activation energy for template religation (41 ± 5 kcal·mol−1) than that for template cleavage (32 ± 1 kcal·mol−1). Our results provide a kinetic interpretation for the ability of Sso topo III to relax negatively supercoiled DNA only at higher temperature and offer clues to the adaptation of the reaction mechanisms of thermophilic enzymes to high temperature.Junhua ZhangBailong PanZhimeng LiXin Sheng ZhaoLi HuangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Junhua Zhang Bailong Pan Zhimeng Li Xin Sheng Zhao Li Huang Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus
description	Abstract All cellular organisms encode type IA topoisomerases which catalyze DNA topological changes essential for DNA transactions. However, the kinetics of the reaction catalyzed by these enzymes remains poorly characterized. Here we measured the rapid kinetics of template binding, cleavage and religation by Sso topo III, a type IA topoisomerase from the hyperthermophilic archaeon Sulfolobus solfataricus, by using a novel FRET/PIFE-based method in a stopped-flow spectrometer. We show that Sso topo III bound the template rapidly, and the rate of binding was 2–3 orders of magnitudes higher than that of template cleavage at 25 °C. The rate of template cleavage was favored over that of template religation by the enzyme, and was more so at lower temperatures (25–55 °C). Significant template cleavage [(2.23 ± 0.11) × 10−3 s−1] was observed while little religation was detectable at 25 °C. This is consistent with the presence of a higher activation energy for template religation (41 ± 5 kcal·mol−1) than that for template cleavage (32 ± 1 kcal·mol−1). Our results provide a kinetic interpretation for the ability of Sso topo III to relax negatively supercoiled DNA only at higher temperature and offer clues to the adaptation of the reaction mechanisms of thermophilic enzymes to high temperature.
format	article
author	Junhua Zhang Bailong Pan Zhimeng Li Xin Sheng Zhao Li Huang
author_facet	Junhua Zhang Bailong Pan Zhimeng Li Xin Sheng Zhao Li Huang
author_sort	Junhua Zhang
title	Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus
title_short	Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus
title_full	Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus
title_fullStr	Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus
title_full_unstemmed	Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus
title_sort	kinetic insights into the temperature dependence of dna strand cleavage and religation by topoisomerase iii from the hyperthermophile sulfolobus solfataricus
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/801302d679f649ae88b2e36adeb5d4dc
work_keys_str_mv	AT junhuazhang kineticinsightsintothetemperaturedependenceofdnastrandcleavageandreligationbytopoisomeraseiiifromthehyperthermophilesulfolobussolfataricus AT bailongpan kineticinsightsintothetemperaturedependenceofdnastrandcleavageandreligationbytopoisomeraseiiifromthehyperthermophilesulfolobussolfataricus AT zhimengli kineticinsightsintothetemperaturedependenceofdnastrandcleavageandreligationbytopoisomeraseiiifromthehyperthermophilesulfolobussolfataricus AT xinshengzhao kineticinsightsintothetemperaturedependenceofdnastrandcleavageandreligationbytopoisomeraseiiifromthehyperthermophilesulfolobussolfataricus AT lihuang kineticinsightsintothetemperaturedependenceofdnastrandcleavageandreligationbytopoisomeraseiiifromthehyperthermophilesulfolobussolfataricus
_version_	1718395054742044672

Kinetic insights into the temperature dependence of DNA strand cleavage and religation by topoisomerase III from the hyperthermophile Sulfolobus solfataricus

Ejemplares similares