title: 5'-methylthioadenosine nucleosidase is implicated in playing a key role in a modified futalosine pathway for menaquinone biosynthesis in Campylobacter jejuni.
abstract: Menaquinone (vitamin K(2)) serves as an electron carrier in the electron transport chain required for respiration in many pathogenic bacteria. Most bacteria utilize a common menaquinone biosynthetic pathway as exemplified by Escherichia coli. Recently, a novel biosynthetic pathway, the futalosine pathway, was discovered in Streptomyces. Bioinformatic analysis strongly suggests that this pathway is also operative in the human pathogens Campylobacter jejuni and Helicobacter pylori. Here, we provide compelling evidence that a modified futalosine pathway is operative in C. jejuni and that it utilizes 6-amino-6-deoxyfutalosine instead of futalosine. A key step in the Streptomyces pathway involves a nucleosidase called futalosine hydrolase. The closest homolog in C. jejuni has been annotated as a 5'-methylthioadenosine nucleosidase (MTAN). We have shown that this C. jejuni enzyme has MTAN activity but negligible futalosine hydrolase activity. However, the C. jejuni MTAN is able to hydrolyze 6-amino-6-deoxyfutalosine at a rate comparable with that of its known substrates. This suggests that the adenine-containing version of futalosine is the true biosynthetic intermediate in this organism. To demonstrate this in vivo, we constructed a C. jejuni mutant strain deleted for mqnA2, which is predicted to encode for the enzyme required to synthesize 6-amino-6-deoxyfutalosine. Growth of this mutant was readily rescued by the addition of 6-amino-6-deoxyfutalosine, but not futalosine. This provides the first direct evidence that a modified futalosine pathway is operative in C. jejuni. It also highlights the tremendous versatility of the C. jejuni MTAN, which plays key roles in S-adenosylmethionine recycling, the biosynthesis of autoinducer molecules, and the biosynthesis of menaquinone.
authors.affiliation: Department of Chemistry, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Xu
authors.initials: X
authors.lastname: Li
authors.forename: Dmitry
authors.initials: D
authors.lastname: Apel
authors.forename: Erin C
authors.initials: EC
authors.lastname: Gaynor
authors.forename: Martin E
authors.initials: ME
authors.lastname: Tanner
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002169:Campylobacter Infections / Q000201:enzymology* / Q000235:genetics; D016123:Campylobacter jejuni / Q000201:enzymology* / Q000235:genetics; D017353:Gene Deletion; D006801:Humans; D009699:N-Glycosyl Hydrolases / Q000235:genetics / Q000378:metabolism*; D009705:Nucleosides / Q000235:genetics / Q000378:metabolism*; D024482:Vitamin K 2 / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D009705:Nucleosides; C122040:futalosine; D024482:Vitamin K 2; D009699:N-Glycosyl Hydrolases