TY - JOUR
T1 - Genetic and biochemical characterizations of aLhr1 helicase in the thermophilic crenarchaeon sulfolobus acidocaldarius
AU - Suzuki, Shoji
AU - Kurosawa, Norio
AU - Yamagami, Takeshi
AU - Matsumoto, Shunsuke
AU - Numata, Tomoyuki
AU - Ishino, Sonoko
AU - Ishino, Yoshizumi
N1 - Funding Information:
Funding: This work was supported by a Grant-in-Aid for Japan Society for the Promotion of Science (JSPS) Fellows (JP20J10898 to S.S.) and by JSPS KAKENHI Grant Number JP19K22289 to Y.I. and JP21K05394 to S.I.
Funding Information:
This work was supported by a Grant-in-Aid for Japan Society for the Promotion of Science (JSPS) Fellows (JP20J10898 to S.S.) and by JSPS KAKENHI Grant Number JP19K22289 to Y.I. and JP21K05394 to S.I. We thank the Center for Advanced instrumental and Educational Supports (Faculty of Agriculture, Kyushu University) for using an image analyzer.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2022/1
Y1 - 2022/1
N2 - Homologous recombination (HR) refers to the process of information exchange between homologous DNA duplexes and is composed of four main steps: end resection, strand invasion and formation of a Holliday junction (HJ), branch migration, and resolution of the HJ. Within each step of HR in Archaea, the helicase-promoting branch migration is not fully understood. Previous biochemical studies identified three candidates for archaeal helicase promoting branch migration in vitro: Hjm/Hel308, PINA, and archaeal long helicase related (aLhr) 2. However, there is no direct evidence of their involvement in HR in vivo. Here, we identified a novel helicase encoded by Saci_0814, isolated from the thermophilic crenarchaeon Sulfolobus acidocaldarius; the helicase dissociated a synthetic HJ. Notably, HR frequency in the Saci_0814-deleted strain was lower than that of the parent strain (5-fold decrease), indicating that Saci_0814 may be involved in HR in vivo. Saci_0814 is classified as an aLhr1 under superfamily 2 helicases; its homologs are conserved among Archaea. Purified protein produced in Escherichia coli showed branch migration activity in vitro. Based on both genetic and biochemical evidence, we suggest that aLhr1 is involved in HR and may function as a branch migration helicase in S. acidocaldarius.
AB - Homologous recombination (HR) refers to the process of information exchange between homologous DNA duplexes and is composed of four main steps: end resection, strand invasion and formation of a Holliday junction (HJ), branch migration, and resolution of the HJ. Within each step of HR in Archaea, the helicase-promoting branch migration is not fully understood. Previous biochemical studies identified three candidates for archaeal helicase promoting branch migration in vitro: Hjm/Hel308, PINA, and archaeal long helicase related (aLhr) 2. However, there is no direct evidence of their involvement in HR in vivo. Here, we identified a novel helicase encoded by Saci_0814, isolated from the thermophilic crenarchaeon Sulfolobus acidocaldarius; the helicase dissociated a synthetic HJ. Notably, HR frequency in the Saci_0814-deleted strain was lower than that of the parent strain (5-fold decrease), indicating that Saci_0814 may be involved in HR in vivo. Saci_0814 is classified as an aLhr1 under superfamily 2 helicases; its homologs are conserved among Archaea. Purified protein produced in Escherichia coli showed branch migration activity in vitro. Based on both genetic and biochemical evidence, we suggest that aLhr1 is involved in HR and may function as a branch migration helicase in S. acidocaldarius.
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U2 - 10.3390/catal12010034
DO - 10.3390/catal12010034
M3 - Article
AN - SCOPUS:85121786119
SN - 2073-4344
VL - 12
JO - Catalysts
JF - Catalysts
IS - 1
M1 - 34
ER -