Helix-hairpin-helix protein MJ1434 from Methanocaldococcus jannaschii and EndoIV homologue TTC0482 from Thermus thermophilus HB27 do not process DNA uracil residues.
The mutagenic threat of hydrolytic DNA cytosine deamination is met mostly by uracil DNA glycosylases (UDG) initiating base excision repair. However, several sequenced genomes of archaeal organisms are devoid of genes coding for homologues of the otherwise ubiquitous UDG superfamily of proteins. Previously, two possible solutions to this problem were offered by (i) a report of a newly discovered family of uracil DNA glycosylases exemplified by MJ1434, a protein found in the hyperthermophilic archaeon Methanocaldococcus jannaschii, and (ii) the description of TTC0482, an EndoIV homologue from the hyperthermophilic bacterium Thermus thermophilus HB27, as being able to excise uracil from DNA. Sequence homologues of both proteins can be found throughout the archaeal domain of life. Three proteins orthologous to MJ1434 and the family founder itself were tested for but failed to exhibit DNA uracil glycosylase activity when produced in an Ung-deficient Escherichia coli host. Likewise, no DNA uracil processing activity could be detected to be associated with TTC0482, while the protein was fully active as an AP endonuclease. We propose that the uracil processing activities formerly found were due to contaminations with Ung enzyme. Use of Deltaung-strains as hosts for production of putatively DNA-U processing enzymes provides a simple safeguard.
Schomacher
Lars
L
Abteilung Molekulare Genetik und Präparative Molekularbiologe, Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Göttingen, Germany.
Smolorz
Sabine
S
Ciirdaeva
Elena
E
Ber
Svetlana
S
Kramer
Wilfried
W
Fritz
Hans-Joachim
HJ
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000378:metabolism; D043223:Deoxyribonuclease IV (Phage T4-Induced) / Q000378:metabolism; D017015:Methanococcales / Q000201:enzymology*; D008969:Molecular Sequence Data; D009154:Mutation; D016415:Sequence Alignment; D016963:Thermus thermophilus / Q000201:enzymology*; D014498:Uracil / Q000378:metabolism; D051981:Uracil-DNA Glycosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D014498:Uracil; D004247:DNA; D043223:Deoxyribonuclease IV (Phage T4-Induced); D051981:Uracil-DNA Glycosidase; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase
Science. 1996 Aug 23;273(5278):1058-73
8688087
Genome Biol. 2000;1(4):RESEARCH0007
11178247
EMBO J. 1996 Oct 1;15(19):5459-69
8895589
Proc Natl Acad Sci U S A. 2003 Nov 11;100(23):13247-52
14585934
J Biol Chem. 1995 Aug 18;270(33):19501-8
7642635
Mol Biol Evol. 2003 Oct;20(10):1603-11
12832627
DNA Repair (Amst). 2004 Nov 2;3(11):1483-92
15380104
J Biol Chem. 1994 Dec 30;269(52):32709-12
7806489
Nature. 1996 Oct 24;383(6602):735-8
8878487
Biochem Biophys Res Commun. 2006 Aug 4;346(3):889-95
16782061
J Bacteriol. 1988 Mar;170(3):1082-91
2963806
J Biol Chem. 1989 Jan 15;264(2):1163-71
2492016
Nucleic Acids Res. 2003 Apr 15;31(8):2045-55
12682355
J Mol Biol. 2002 Jan 18;315(3):373-84
11786018
Biochemistry. 1990 Mar 13;29(10):2532-7
2185829
Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1642-6
8127859
J Biol Chem. 1999 Mar 12;274(11):7128-36
10066771
Nucleic Acids Res. 2002 May 15;30(10):2097-102
12000829
Proc Natl Acad Sci U S A. 2002 Apr 2;99(7):4644-9
11930014
J Bacteriol. 2004 Oct;186(20):6956-69
15466049
Annu Rev Biochem. 1994;63:915-48
7979257
Biochemistry. 2000 Aug 22;39(33):10098-109
10955998
J Bacteriol. 2006 Jan;188(2):642-58
16385054
J Bacteriol. 1997 Nov;179(22):7135-55
9371463
J Biol Chem. 1977 May 25;252(10):3286-94
324994
Curr Biol. 1999 Feb 25;9(4):174-85
10074426
Nucleic Acids Res. 2006;34(18):5325-36
17012282
DNA Repair (Amst). 2010 Apr 4;9(4):438-47
20129830
Curr Biol. 1999 May 20;9(10):531-4
10339434
Biochemistry. 1987 Dec 15;26(25):8200-6
3327518
EMBO J. 1995 Aug 15;14(16):4108-20
7664751
Mikrobiologiia. 2008 Sep-Oct;77(5):644-50
19004346
Protein Eng. 1999 Apr;12(4):333-40
10325404
J Biol Chem. 1977 May 10;252(9):2808-14
16002
Nucleic Acids Res. 2009 Apr;37(7):2283-93
19240141
Proc Natl Acad Sci U S A. 2004 Apr 27;101(17):6391-6
15096615
Biochem J. 1997 Jul 1;325 ( Pt 1):1-16
9224623
Biotechniques. 1992 Aug;13(2):180-4
1389144
Nature. 1993 Apr 22;362(6422):709-15
8469282
Curr Biol. 1999 Jun 17;9(12):653-6
10375529