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Micrtxilnns International 300 N.Zeeb Road Ann Arbor, Ml 48106 INFORMATION TO USERS This reproduction was made from a copy of a document sent to us for microfilming. While the most advanced technology has been used to photograph and reproduce this document, the quality of the reproduction is heavily dependent upon the quality of the material submitted. The following explanation of techniques is provided to help clarify markings or notations which may appear on this reproduction. 1. The sign or “target” for pages apparently lacking from the document photographed is “Missing Page(s)”. If it was possible to obtain the missing page(s) or section, they are spliced into the film along with adjacent pages. This may have necessitated cutting through an image and duplicating adjacent pages to assure complete continuity. 2. 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These prints are available upon request from the Dissertations Customer Services Department. 5. Some pages in any document may have indistinct print. In aU cases the best available copy has been filmed. U niversi^ Micrtxilnns International 300 N.Zeeb Road Ann Arbor, Ml 48106 8426494 T ry o n , V icto r V. PURINE SALVAGE METABOLISM IN MOLLICUTES The Ohio State University Ph.D. 1984 University Microfilms Internstionel300 N. Zeeb Road, Ann Arbor. Ml48106 Copyright 1984 by Tryon, Victor V. All Rights Reserved PLEASE NOTE: In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a check mark V_ 1. Glossy photographs or pages. 2. Colored illustrations, paper or print. 3. Photographs with dark background _____ 4. Illustrations are poor copy ______ 5. Pages with black marks, not original copy. 6. Print shows through as there is text on both sides of page. 7. Indistinct, broken or small print on several pages _ 8. Print exceeds margin requirements _____ 9. Tightly bound copy with print lost in spine ______ 10. Computer printout pages with indistinct print. 11. Page(s) ___________ lacking when material received, and not available from school or author. 12. Page(s)____________seem to be missing in numbering only as text follows. 13. Two pages numbered ___________ . Text follows. 14. Curling and wrinkled pages ______ 15. Other__________________________________________________________________ University Microfilms International PURINE SALVAGE METABOLISM IN MOLLICUTES DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio S tate University By Victor V. Tryon, 8.S. ***** The Ohio State University 1984 Reading Committee: Approved By: J . Dennis Pollack, Ph.D. Frank A. Kapral, Ph.D. Abramo C. Ottolenghi, Ph.D. Adviser Norman L. Somerson, Ph.D. ïépartment of Medical Microbiology Marshall V. Williams, Ph.D. and Immunology Copyright by Victor V. Tryon 1984 To Nancy n I wish to thank Dr. J. D. Pollack for his considerable efforts on my behalf. Dr. M. V. Williams was of great assistance with the enzyme work. I should also like to thank Drs. F. A. Kapral, A. 0. Ottolenghi and N. L. Somerson for their advice and encourage­ ment. R. Montione provided special help with the graphic photography. m VITA November 14, 1953 ................. Born - Oakland, C alifo rn ia 1981 ........................................... B.S., University of Washington Seattle, Washington 1981-1982 .................................. Graduate Student, Department of Medical M icrobiology and Immunology, The Ohio State University, Columbus, Ohio 1982-1984 .................................. Research Associate, Department of Medical M icrobiology and Iiranunology, The Ohio State University, Columbus, Ohio PUBLICATIONS P ollack, J .D ., V.V. Tryon, and K.D. Beaman. 1983. The m etabolic pathways of Acholeplasma and Mycoplasma: An overview. Yale 0. Med. 56:709-7T6. Tryon, V.V., and J.D. Pollack. 1984 Purine metabolism in Achole­ plasma laidlawii B-PG9: A novel pyrophosphate dependent nucleoside kinase activity. J. Bacteriol. 159:265-270. ABSTRACTS AND PRESENTATIONS P ollack, J .D ., Beaman, K.D., Tryon, V.V., and Robertson, J . New emerging patterns of NADH oxidase localization and lipid synthesis in Mollicutes. Int. Org. Mycoplasmology, Tokyo, Japan, September, 1982. Tryon, V.V., and J.D. Pollack. Purine salvage is the sole route for the synthesis of cellular adenylates in Acholeplasma laidlawii B-PG9. American S ociety fo r M icrobiology. New O rleans, LA, March 6, 1983. Tryon, V.V., and J.D. Pollack. Purine salvage and interconversion in Acholeplasma laidlawii B-PG9. American Society for Micro- biology. St. Louis, MO, March 9, 1984. Pollack, J.D., V.V. Tryon, and M.V. Williams. Pyrophosphate metabolism in A. la id la w ii B-PG9. F ifth In te rn a tio n a l Congress of the I n te r ­ national Organization of Mycoplasmology. Jerusalem, Israel, June, 1984. iv VITA, co n tin u ed FIELD OF STUDY Major Field: Medical Microbiology and Immunology Studies in Microbial Physiology and Enzymology: Professor J. Dennis Pollack Studies in Mycoplasmology: Professor J. Dennis Pollack TABLE OF CONTENTS Page DEDICATION ............................................................................................ ü ACKNOWLEDGEMENTS ................................................................................. i i i VITA .................................................................................................... iv LIST OF TABLES..................................................................................... vii LIST OF FIGURES ................................................................................. v iii LIST OF PLATES..................................................................................... ix INTRODUCTION ......................................................................................... 1 MATERIALS AND METHODS ...................................................................... 8 RESULTS................................................................................................ 19 DISCUSSION............................................................................................. 32 CONCLUSIONS......................................................................................... 45 LITERATURE CITED................................................................................ 47 VI LIST OF TABLES Table Page 1 Organisms used in th is study ........................................... 9 2 Summary of purine enzymes assayed ............................... 16,17 3 Purine salvage enzymes in Acholeplasma laidlawii B-PG9 ............................................................................. 22 4 Purine salvage enzymes in Acholeplasma axanthum S-743 ...............................................■■............................ 23 5 Purine salvage enyzmes in Acholeplasma granularum BTS-39 .......................................................................... 24 6 Purine salvage enzymes in Mycoplasme gallisepticum S 6 ................................................................................. 225 7 Purine salvage enzymes in Spiroplasma floricola 236. 26 8 Purine salvage enzymes in Mycoplasma arqinini G-230. 27 9 Purine salvage enzymes in Acholeplasma florum LI . 28 10 Nucleoside kinase activities in A. laidlawii B-PG9, A. axanthum S-743, Acholeplasma granularum BTS-39 and coli (ATCC 25922) ........................... 29 11 Summary of purine salvage enzyme activities .... 30 12 Known reactions in which PPi replaces ATP as a phosphate donor .......................................................... 40 vn LIST OF FIGURES Figure Figure Legends Page 1 Effect of various concentrations of sodium pyrophosphate or ATP on the amount of AMP synthesized in the standard reaction mixture for adenosine kinase activity by dialyzed lysates of Acholeplasma laidlaw ii B-P69 ................ 31 2 Proposed salvage pathways for the biosynthe­ sis of purine nucleotides in Acholeplasma laidlaw ii B-PG9 .............................................................. 33 3 Recovery of labeled AMP, adenosine and adenine during the assay of adenine phosphoribosyl- transferase activity in Acholeplasma laidlawii B-PG9 with respect to time ....................................... 36 4 Proposed salvage pathways for the biosynthesis of purine nucleotides in Acholeplasma florum LI and Spiroplasma floricola ........................................... 43 5 Proposed salvage pathways for the biosynthesis of purine nucleotides in Mycoplasma gal1isepticum and Mycoplasma arqinini ............................................... 44 v m LIST OF PLATES PI ate Page A Autoradiograph
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