Co-expression with RadA and the characterization of stRad55B, a RadA paralog from the hyperthermophilic crenarchaea Sulfolobus tokodaii.
ST0838 (designed stRad55B) is one of the four RadA paralogs (or Rad55 homologues) in the genome of the hyperthermophilic crenarchaeon Sulfolobus tokodaii. The gene is induced by UV irradiation, suggesting that it is involved in DNA recombinational repair in this organism. However, this protein could not be expressed normally in vitro. In this study, thermostable and soluble stRad55B was obtained by co-expression with S. tokodaii RadA (stRadA) in E. coli, and the enzymatic properties were examined. It was found that stRad55B bound ssDNA preferentially and had a very weak ATPase activity that was not stimulated by DNA. The recombinant protein inhibited the strand exchange activity promoted by stRadA, indicating that stRad55B might be an inhibitor to the homologous recombination in this archaeon. The results will be helpful for further functional and interaction analysis of RadA paralogs and for the understanding of the mechanism of recombinational repair in archaea.
Sheng
DuoHong
D
State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China.
Li
MingFeng
M
Jiao
JianDong
J
Ni
JinFeng
J
Shen
YuLong
Y
D000251:Adenosine Triphosphatases / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000528:radiation effects; D011485:Protein Binding; D011489:Protein Denaturation; D016944:Sulfolobus / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D013696:Temperature
D019843:Archaeal Proteins; D004268:DNA-Binding Proteins; D004247:DNA; D000251:Adenosine Triphosphatases