Preferences help
enabled [disable] Abstract
Number of results
2014 | 61 | 2 | 393-395
Article title

The pH optimum of native uracil-DNA glycosylase of Archaeoglobus fulgidus compared to recombinant enzyme indicates adaption to cytosolic pH

Title variants
Languages of publication
Uracil-DNA glycosylase of Archaeoglobus fulgidus (Afung) in cell extracts exhibited maximal activity around pH 6.2 as compared to pH 4.8 for the purified recombinant enzyme expressed in Escherichia coli. Native Afung thus seems to be adapted to the intracellular pH of A. fulgidus, determined to be 7.0±0.1. Both recombinant and native Afung exhibited a broad temperature optimum for activity around 80°C, reflecting the A. fulgidus optimal growth temperature of 83°C. Adaption to the neutral conditions in the A. fulgidus cytoplasm might be due to covalent modifications or accessory factors, or due to a different folding when expressed in the native host.
Physical description
  • Faculty of Science and Technology, Department of Mathematics and Natural Sciences-Centre for Organelle Research, University of Stavanger, Stavanger, Norway
  • Faculty of Science and Technology, Department of Mathematics and Natural Sciences-Centre for Organelle Research, University of Stavanger, Stavanger, Norway
  • Department of Biology and Centre for Geobiology, University of Bergen, Bergen, Norway
  • Faculty of Science and Technology, Department of Mathematics and Natural Sciences-Centre for Organelle Research, University of Stavanger, Stavanger, Norway
  • Behzadi A, Hatleskog R, Ruoff P (1999) Hysteretic enzyme adaptation to environmental pH: Change in storage pH of alkaline phosphatase leads to a pH-optimum in the opposite direction to the applied change. Biophys Chem 77: 99-109.
  • Bjelland S, Seeberg E (1987) Purification and characterization of 3-methyladenine DNA glycosylase I from Escherichia coli. Nucleic Acids Res 15: 2787-2801.
  • Boal AK, Yavin E, Lukianova OA, O'Shea VL, David SS, Barton JK (2005) DNA-bound redox activity of DNA repair glycosylases containing [4Fe-4S] clusters. Biochemistry 44: 8397-8407.
  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.
  • Chung JH, Im EK, Park HY, Kwon JH, Lee S, Oh J, Hwang KC, Lee JH, Jang Y (2003) A novel uracil-DNA glycosylase family related to the helix-hairpin-helix DNA glycosylase superfamily. Nucleic Acids Res 31: 2045-2055.
  • Engstrom LM, Partington OA, David SS (2012) An iron-sulfur cluster loop motif in the Archaeoglobus fulgidus uracil-DNA glycosylase mediates efficient uracil recognition and removal. Biochemistry 51: 5187-5197.
  • Frederico LA, Kunkel TA, Shaw BR (1990) A sensitive genetic assay for the detection of cytosine deamination: Determination of rate constants and the activation energy. Biochemistry 29: 2532-2537.
  • Grogan DW (2004) Stability and repair of DNA in hyperthermophilic archaea. Curr Issues Mol Biol 6: 137-144.
  • Hagen L, Kavli B, Sousa MM, Torseth K, Liabakk NB, Sundheim O, Peňa-Diaz J, Otterlei M, Hørning O, Jensen ON, Krokan HE, Slupphaug G (2008) Cell cycle-specific UNG2 phosphorylations regulate protein turnover, activity and association with RPA. EMBO J 27: 51-61.
  • Johnson WC, Lindsey AJ (1939) An improved universal buffer. Analyst (London) 64: 490-492.
  • Knævelsrud I, Moen MN, Grøsvik K, Haugland GT, Birkeland NK, Klungland A, Leiros I, Bjelland S (2010) The hyperthermophilic euryarchaeon Archaeoglobus fulgidus repairs uracil by single-nucleotide replacement. J Bacteriol 192: 5755-5766.
  • Knævelsrud I, Ruoff P, Ånensen H, Klungland A, Bjelland S, Birkeland NK (2001) Excision of uracil from DNA by the hyperthermophilic Afung protein is dependent on the opposite base and stimulated by heat-induced transition to a more open structure. Mutat Res 487: 173-190.
  • Kornberg A, Baker TA (1992) DNA Replication, 2nd edition. New York: W.H. Freeman.
  • Koulis A, Cowan DA, Pearl LH, Savva R (1996) Uracil-DNA glycosylase activities in hyperthermophilic micro-organisms. FEMS Microbiol Lett 143: 267-271.
  • Krokan HE, Standal R, Slupphaug G (1997) DNA glycosylases in the base excision repair of DNA. Biochem J 325: 1-16.
  • LaRonde-LeBlanc N, Guszczynski T, Copeland T, Wlodawer A (2005a) Autophosphorylation of Archaeoglobus fulgidus Rio2 and crystal structures of its nucleotide-metal ion complexes. FEBS J 272: 2800-2810.
  • LaRonde-LeBlanc N, Guszczynski T, Copeland T, Wlodawer A (2005b) Structure and activity of the atypical serine kinase Rio1. FEBS J 272: 3698-3713.
  • Lindahl T (1974) An N-glycosidase from Escherichia coli that releases free uracil from DNA containing deaminated cytosine residues. Proc Natl Acad Sci USA 71: 3649-3653.
  • Lindahl T (1993) Instability and decay of the primary structure of DNA. Nature 362: 709-715.
  • Lindahl T, Nyberg B (1974) Heat-induced deamination of cytosine residues in deoxyribonucleic acid. Biochemistry 13: 3405-3410.
  • Lund P (2011) Insights into chaperonin function from studies on archaeal thermosomes. Biochem Soc Trans 39: 94-98.
  • Pearl LH (2000) Structure and function in the uracil-DNA glycosylase superfamily. Mutat Res 460: 165-181.
  • Sandigursky M, Franklin WA (2000) Uracil-DNA glycosylase in the extreme thermophile Archaeoglobus fulgidus. J Biol Chem 275: 19146-19149.
  • Sartori AA, Fitz-Gibbon S, Yang H, Miller JH, Jiricny J (2002) A novel uracil-DNA glycosylase with broad substrate specificity and an unusual active site. EMBO J 21: 3182-3191.
  • Shi L, Potts M, Kennelly PJ (1998) The serine, threonine, and/or tyrosine-specific protein kinases and protein phosphatases of prokaryotic organisms: A family portrait. FEMS Microbiol Rev 22: 229-253.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.