Graduate Thesis Or Dissertation
 

A quantitative comparison of the incorporation of C¹⁴ labeled lipids and lipid precursors into Lee influenza virus

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  • A quantitative comparison was made of the incorporation of 14, C¹⁴ lipid compounds into Lee influenza virus. The compounds tested were either lipids reported to be present in the virus or precursors to virus lipid components. Infected chorioallantoic tissues were incubated in medium containing the C¹⁴ metabolites. The virus was purified by specific adsorption to chicken red blood cells, ultracentrifugation and chromatography on an ECTEOLA anion exchange column. The purified virus was titrated and the C¹⁴measured by scintillation counting. The C¹⁴ in the tissues was also determined. The incorporation of the compounds into the infected tissues was measured and compared to the uptake by normal tissues. All but two of the compounds tested were incorporated into the virus in significant amounts. Ethanolamine and choline, precursors to phospholipids found in the virus, were not incorporated into the virus. This suggested the major virus phospholipids were synthesized before infection. However, glycerol, another phospholipid precursor, was utilized in virus production. It is possible that some of the minor phosphatides of the virus, such as phosphatidic acid, were synthesized after infection. It appeared that there was a significant decrease in fatty acid synthesis after infection. Only a small amount of malonic acid was incorporated into the virus. Due to their solubility, the fatty acids supplied to the infected host cells were divided into two groups. The long chain acids did not seem to be utilized to a great extent for virus formation. The short chain acids contributed significantly greater amounts of label to the virus, in terms of molar quantities, than any of the long chain acids. The value for butyric acid was greater than for any other fatty acid. In addition, this fatty acid stimulated virus production nearly two fold. The results suggested that most of the virus cholesterol was preformed host material. There was little incorporation into the virus and infected tissue from the precursor 3-hydroxy-3-methyl-glutaric acid-3-C¹⁴. The infected tissue incorporated significant amounts of C¹⁴ from all of the compounds studied. There seemed to be a relation between the amount of C¹⁴ incorporated into the tissue and that found in the virus, except in the case of ethanolamine and choline. Both of these compounds were found in the infected tissues in substantial amounts but were not utilized in new virus formation. Results indicated that eight of the fourteen compounds tested were incorporated significantly less in infected tissue than in control tissue.
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