Studies on the ADP-ribose pyrophosphatase subfamily of the nudix hydrolases and tentative identification of trgB, a gene associated with tellurite resistance.
Four Nudix hydrolase genes, ysa1 from Saccharomyces cerevisiae, orf209 from Escherichia coli, yqkg from Bacillus subtilis, and hi0398 from Hemophilus influenzae were amplified, cloned into an expression vector, and transformed into E. coli. The expressed proteins were purified and shown to belong to a subfamily of Nudix hydrolases active on ADP-ribose. Comparison with other members of the subfamily revealed a conserved proline 16 amino acid residues downstream of the Nudix box, common to all of the ADP-ribose pyrophosphatase subfamily. In this same region, a conserved tyrosine designates another subfamily, the diadenosine polyphosphate pyrophosphatases, while an array of eight conserved amino acids is indicative of the NADH pyrophosphatases. On the basis of these classifications, the trgB gene, a tellurite resistance factor from Rhodobacter sphaeroides, was predicted to designate an ADP-ribose pyrophosphatase. In support of this hypothesis, a highly specific ADP-ribose pyrophosphatase gene from the archaebacterium, Methanococcus jannaschii, introduced into E. coli, increased the transformant's tolerance to potassium tellurite.
Dunn
C A
CA
Department of Biology and the McCollum-Pratt Institute, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
O'Handley
S F
SF
Frick
D N
DN
Bessman
M J
MJ
D000251:Adenosine Triphosphatases / Q000633:toxicity; D000595:Amino Acid Sequence; D000818:Animals; D053502:Arsenate Reductases; D053501:Arsenite Transporting ATPases; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D004351:Drug Resistance; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D015723:Gene Library; D006801:Humans; D016623:Ion Pumps; D017017:Methanococcus / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D011755:Pyrophosphatases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D012242:Rhodobacter sphaeroides / Q000201:enzymology / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D013691:Tellurium / Q000494:pharmacology*
D016623:Ion Pumps; D009097:Multienzyme Complexes; D029701:Saccharomyces cerevisiae Proteins; C506971:ARR2 protein, S cerevisiae; D053502:Arsenate Reductases; C058796:ADP-ribose pyrophosphatase I; C058797:ADP-ribose pyrophosphatase II; D000251:Adenosine Triphosphatases; D011755:Pyrophosphatases; D053501:Arsenite Transporting ATPases; C026660:tellurous acid; D013691:Tellurium