Endotoxicosis Induced by Coxiella burnetii Lipopolysaccharide Stimulates a Ribosomal Protein S6 Kinase: Some Properties of the Partially Purified Enzyme

Guinea pig endotoxicosis induced by lipopolysaccharide from Coxiella burnetii Nine Mile phase I stimulates phosphorylation of liver ribosomal protein S6, with a 50% increase at 12 h postinoculation. The responsible protein kinase (S6PK) has been partially purified from liver; its activity is independent of cyclic AMP and of Ca2+ plus phosphatidyl serine or diacylglycerol. The preparation has an apparent optimum concentration of 20 mM Mg2+, while Ca2+ and Mn2+ are each inhibitory at 2 mM. The apparent Km for ATP is 30 microM with intact ribosomes. Because of the central role of phosphorylation in metabolic regulation and a purported role of phosphorylated S6 in protein synthesis, the lipopolysaccharide-induced stimulation of S6PK suggests a significant regulatory role of such enzymes in the pathobiochemistry of Q fever infection and endotoxicosis

Changes in Hepatic Glycogen, Protein, and Ribonucleic Acid Synthesis, and Some Effects of Cortisol, During Q Fever

Liver glycogen is depleted in guinea pigs infected with Coxiella burneti . Syntheses of the glycogen precursors uridine triphosphate and uridine diphosphate glucose are unaffected during Q fever, but glycogen synthetase activity is inhibited. Exogenous cortisol relieves this inhibition in infected animals. Orotate and amino acids are more rapidly incorporated into ribonucleic acid and protein during infection. It is proposed that the biochemical defect in the synthesis of glycogen lies in the inactivation of glycogen synthetase. </jats:p

STUDIES ON THE PHYSIOLOGY OF RICKETTSIAE V

Mallavia , L. (University of Kansas, Lawrence) and D. Paretsky . Studies on the physiology of rickettsiae. V. Metabolism of carbamyl phosphate by Coxiella burnetii . J. Bacteriol. 86: 232–238. 1963.—Preparations of disrupted Coxiella burnetii catalyze synthesis of citrulline from ornithine and carbamyl phosphate at an optimal pH of 7.0 to 7.5. Rickettsial synthesis of the pyrimidine precursor, ureidosuccinate, is demonstrated and confirmed by isolating C 14 -labeled ureidosuccinate from reaction mixtures of carbamyl phosphate and labeled aspartate. The data suggest a further rickettsial synthesis of orotate and imply rickettsial competence for host-independent pyrimidine synthesis. </jats:p

Chemical and Biochemical Changes in Subcellular Fractions of Guinea Pig Liver During Infection with <i>Coxiella burneti</i>

Livers of uninfected guinea pigs and of guinea pigs infected with Coxiella burneti were fractionated into smooth endoplasmic reticulum, rough endoplasmic reticulum (RER), pellet, and cell sap fractions. The ribonucleic acid (RNA) and protein of each fraction were determined, and the phosphorylase, glucose-6-phosphatase, and glucosyl transferase (glycogen synthetase) activities of each fraction were measured. Decreased RNA, protein, and enzyme activities were found in the RER and pellet fractions of infected livers, with the greatest differences in the RER. The evidence indicates a solubilization of the phosphorylase and synthetase, with the enzymes moving from the RER and glycogen-containing pellet fraction to the cell sap. The data suggest the RER as a target during Q fever. </jats:p