Efficiency of purine utilization by Helicobacter pylori: roles for adenosine deaminase and a NupC homolog.
The ability to synthesize and salvage purines is crucial for colonization by a variety of human bacterial pathogens. Helicobacter pylori colonizes the gastric epithelium of humans, yet its specific purine requirements are poorly understood, and the transport mechanisms underlying purine uptake remain unknown. Using a fully defined synthetic growth medium, we determined that H. pylori 26695 possesses a complete salvage pathway that allows for growth on any biological purine nucleobase or nucleoside with the exception of xanthosine. Doubling times in this medium varied between 7 and 14 hours depending on the purine source, with hypoxanthine, inosine and adenosine representing the purines utilized most efficiently for growth. The ability to grow on adenine or adenosine was studied using enzyme assays, revealing deamination of adenosine but not adenine by H. pylori 26695 cell lysates. Using mutant analysis we show that a strain lacking the gene encoding a NupC homolog (HP1180) was growth-retarded in a defined medium supplemented with certain purines. This strain was attenuated for uptake of radiolabeled adenosine, guanosine, and inosine, showing a role for this transporter in uptake of purine nucleosides. Deletion of the GMP biosynthesis gene guaA had no discernible effect on mouse stomach colonization, in contrast to findings in numerous bacterial pathogens. In this study we define a more comprehensive model for purine acquisition and salvage in H. pylori that includes purine uptake by a NupC homolog and catabolism of adenosine via adenosine deaminase.
Microbiology Department, The University of Georgia, Athens, Georgia, United States of America.
Erica F
EF
Miller
Soumya
S
Vaish
Robert J
RJ
Maier
D000241:Adenosine / Q000378:metabolism; D000243:Adenosine Deaminase / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D018929:Cell Culture Techniques; D003470:Culture Media; D005260:Female; D005753:Gastric Mucosa / Q000382:microbiology*; D016480:Helicobacter pylori / Q000254:growth & development* / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D008969:Molecular Sequence Data; D011687:Purines / Q000378:metabolism*; D016415:Sequence Alignment; D013045:Species Specificity
D001426:Bacterial Proteins; D003470:Culture Media; D026901:Membrane Transport Proteins; C088081:NupC protein, Bacteria; D011687:Purines; D000243:Adenosine Deaminase; D000241:Adenosine