Enzymatic activities leading to pyrimidine nucleotide biosynthesis from cell-free extracts of Rickettsia typhi.
Cell-free extracts from Rickettsia typhi were examined for the presence or absence of pyrimidine phosphotransferase enzymes and compared with the enzymes of mouse L cells and Salmonella typhimurium. The organisms were grown in mouse L cells and in the yolk sacs of chicken embryos, purified by Renografin density gradient centrifugation, and ruptured in a French pressure cell. The enzymes for the reutilization of uridine and thymidine, uridine kinase (EC 2.7.1.48) and thymidine kinase (EC 2.7.1.21), were not detected in R. typhi extracts with the phosphate donors effective for control enzymes. The following enzyme activities were demonstrated in R. typhi: uridine-5'-monophosphate kinase (UMPK, EC  2.7.4.4), deoxythymidine-5'-monophosphate kinase (dTMPK, EC 2.7.4.9), and nucleosidediphosphate kinase (NDPK, EC 2.7.4.6). Physicochemical and enzymatic analyses demonstrated that the pyrimidine nucleotide kinases of R. typhi were not of host origin and that the source (yolk sac and mouse L cells) did not influence the relative enzymatic activities. The specific activities of UMPK and dTMPK were higher when the rickettsiae were harvested before embryo death, whereas NDPK levels were slightly decreased. The specific activities of UMPK, dTMPK, and NDPK were comparable to those of S. typhimurium, and consequently the rickettsiae have potential for the anabolism of monophosphates, as do the host-independent bacteria. These results suggest that R. typhi cannot utilize host uridine or thymidine pools directly but must rely on themonophosphorylated molecules of the host cell or must synthesize the monophosphates de novo.
D000818:Animals; D002474:Cell-Free System; D002642:Chick Embryo; D005260:Female; D007739:L Cells / Q000201:enzymology; D051379:Mice; D010770:Phosphotransferases / Q000378:metabolism; D011742:Pyrimidine Nucleotides / Q000096:biosynthesis*; D012286:Rickettsia typhi / Q000201:enzymology*; D012486:Salmonella typhimurium / Q000201:enzymology; D013937:Thymidine Kinase / Q000378:metabolism; D013997:Time Factors; D014541:Uridine Kinase / Q000378:metabolism; D014817:Vitelline Membrane / Q000201:enzymology / Q000382:microbiology
D011742:Pyrimidine Nucleotides; D010770:Phosphotransferases; D013937:Thymidine Kinase; D014541:Uridine Kinase