Helicobacter pylori extracts exhibit nicotinamide adenine dinucleotide-derived adenylation but not mono(adenosine 5'-diphosphate-ribosyl)ation of DNA ligase.
The issue of toxins produced by Helicobacter pylori (H. pylori) urgently requires clarification given that the bacterium causes gastric epithelial cell damage which may lead to precancerous and cancerous changes. During an investigation of the possibility of mono(adenosine 5'-diphosphate (ADP)-ribosyl)ation by H. pylori products, as observed for other bacterial toxins, we found that radioactivity of [adenylate-32P]nicotinamide adenine dinucleotide (NAD) is incorporated into an H. pylori protein of 80 kDa after incubation with crude bacterial extract. In contrast, [carbonyl-14C]NAD did not show any radioactivity incorporation. Unexpectedly, treatment of the modified protein with 0.1 N HCl, but not 0.1 N NaOH, released the AMP moiety. Such chemical properties are characteristic of bacterial DNA ligase-AMP complexes. We found that an antibody raised against Escherichia coli DNA ligase [EC 6.5.1.2] immunoprecipitated the modified 80 kDa protein. Our results indicate that incorporation of radioactivity derived from NAD into the 80 kDa protein was due to adenylation, but not mono(ADP-ribosyl)ation, of the DNA ligase of H. pylori.
Nozaki
T
T
Biochemistry Division, National Cancer Center Research Institute.
Masutani
M
M
Noda
T
T
Saito
D
D
Sugiyama
T
T
Takato
T
T
Wakabayashi
K
K
Nakagama
H
H
Sugimura
T
T
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D001426:Bacterial Proteins / Q000276:immunology; D011088:DNA Ligases / Q000276:immunology / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D016480:Helicobacter pylori / Q000302:isolation & purification / Q000378:metabolism*; D006851:Hydrochloric Acid / Q000494:pharmacology; D009243:NAD / Q000378:metabolism*; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D011233:Precipitin Tests; D012263:Ribonucleosides / Q000032:analysis*; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000494:pharmacology; D012972:Sodium Hydroxide / Q000494:pharmacology; D013820:Thermolysin / Q000494:pharmacology
D001426:Bacterial Proteins; D009702:Nucleoside Diphosphate Sugars; D012263:Ribonucleosides; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D012972:Sodium Hydroxide; D015719:Single-Strand Specific DNA and RNA Endonucleases; D013820:Thermolysin; D011088:DNA Ligases; D006851:Hydrochloric Acid