title: Rickettsial metK-encoded methionine adenosyltransferase expression in an Escherichia coli metK deletion strain.
abstract: The obligate intracellular bacterium Rickettsia prowazekii has recently been shown to transport the essential metabolite S-adenosylmethionine (SAM). The existence of such a transporter would suggest that the metK gene, coding for the enzyme that synthesizes SAM, is unnecessary for rickettsial growth. Genome sequencing has revealed that this is the case for the metK genes of the spotted fever group and the Madrid E strain of R. prowazekii, which contain recognizable inactivating mutations. However, several strains of the typhus group rickettsiae possess metK genes lacking obvious mutations. In order to determine if these genes code for a product that retains MAT function, an Escherichia coli metK deletion mutant was constructed in which individual rickettsial metK genes were tested for the ability to complement the methionine adenosyltransferase deficiency. Both the R. prowazekii Breinl and R. typhi Wilmington metK genes complemented at a level comparable to that of an E. coli metK control, demonstrating that the typhus group rickettsiae have the capability of synthesizing as well as transporting SAM. However, the appearance of mutations that affect the function of the metK gene products (a stop codon in the Madrid E strain and a 6-bp deletion in the Breinl strain) provides experimental support for the hypothesis that these typhus group genes, like the more degenerate spotted fever group orthologs, are in the process of gene degradation.
authors.affiliation: Department of Microbiology and Immunology, Laboratory of Molecular Biology, University of South Alabama, Mobile, AL 36688-0002, USA.
authors.forename: Lonnie O
authors.initials: LO
authors.lastname: Driskell
authors.forename: Aimee M
authors.initials: AM
authors.lastname: Tucker
authors.forename: Herbert H
authors.initials: HH
authors.lastname: Winkler
authors.forename: David O
authors.initials: DO
authors.lastname: Wood
mesh_terms: D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D008716:Methionine Adenosyltransferase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D012283:Rickettsia prowazekii / Q000201:enzymology / Q000235:genetics*; D012286:Rickettsia typhi / Q000201:enzymology / Q000235:genetics*; D012436:S-Adenosylmethionine / Q000378:metabolism*
chemical_list: D012436:S-Adenosylmethionine; D008716:Methionine Adenosyltransferase