title: Bacillus subtilis 5'-nucleotidases with various functions and substrate specificities.
abstract: In Escherichia coli, nagD, yrfG, yjjG, yieH, yigL, surE, and yfbR encode 5'-nucleotidases that hydrolyze the phosphate group of 5'-nucleotides. In Bacillus subtilis, genes encoding 5'-nucleotidase have remained to be identified.
  
  
  
  We found that B. subtilis ycsE, araL, yutF, ysaA, and yqeG show suggestive similarities to nagD. Here, we expressed them in E. coli to purify the respective His6-tagged proteins. YcsE exhibited significant 5'-nucleotidase activity with a broader specificity, whereas the other four enzymes had rather weak but suggestive activities with various capacities and substrate specificities. In contrast, B. subtilis yktC shares high similarity with E. coli suhB encoding an inositol monophosphatase. YktC exhibited inositol monophosphatase activity as well as 5'-nucleotidase activity preferential for GMP and IMP. The ycsE, yktC, and yqeG genes are induced by oxidative stress and were dispensable, although yqeG was required to maintain normal growth on solid medium. In the presence of diamide, only mutants lacking yktC exhibited enhanced growth defects, whereas the other mutants without ycsE or yqeG did not.
  
  
  
  Accordingly, in B. subtilis, at least YcsE and YktC acted as major 5'-nucleotidases and the four minor enzymes might function when the intracellular concentrations of substrates are sufficiently high. In addition, YktC is involved in resistance to oxidative stress caused by diamide, while YqeG is necessary for normal colony formation on solid medium.
authors: Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Ayako
authors: A
authors: Terakawa
authors: Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Ayane
authors: A
authors: Natsume
authors: Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Atsushi
authors: A
authors: Okada
authors: Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Shogo
authors: S
authors: Nishihata
authors: Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Junko
authors: J
authors: Kuse
authors: Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Kosei
authors: K
authors: Tanaka
authors: Department of Agrobioscience, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.|Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Shinji
authors: S
authors: Takenaka
authors: Department of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan.
authors: Shu
authors: S
authors: Ishikawa
authors: Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan. kenyoshi@kobe-u.ac.jp.|Department of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo, 657-8501, Japan. kenyoshi@kobe-u.ac.jp.
authors: Ken-Ichi
authors: KI
authors: Yoshida
mesh_terms: D015720:5'-Nucleotidase / Q000235:genetics / Q000378:metabolism*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins / Q000378:metabolism; D005798:Genes, Bacterial; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D007295:Inositol Phosphates / Q000378:metabolism; D009708:Nucleotidases / Q000378:metabolism; D018384:Oxidative Stress / Q000235:genetics; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism; D017385:Sequence Homology; D013234:Stem Cells; D013379:Substrate Specificity
chemical_list: D029968:Escherichia coli Proteins; D007295:Inositol Phosphates; D006867:Hydrolases; C509417:NagD protein, E coli; D009708:Nucleotidases; D010744:Phosphoric Monoester Hydrolases; C021822:myo-inositol-1 (or 4)-monophosphatase; D015720:5'-Nucleotidase; C034507:2-haloacid dehalogenase
keywords: 5′-nucleotidase; Bacillus subtilis; Haloacid dehalogenase superfamily; Inositol monophosphatase; Inositol phosphate; Nucleoside/nucleotide metabolism; Oxidative stress; Phosphatase; Protein motif