Synthesis of FUDP-N-acetylglucosamine and FUDP-glucose in Saccharomyces cerevisiae cells treated with 5-fluorouracil.
Saccharomyces cerevisiae cells (strain W303-1A) treated with 5-fluorouracil and grown in 2% (fermentative conditions) or in 0.1% glucose (oxidative conditions) accumulated two types of 5-fluoro-UDP-sugars (FUDP-sugars): FUDP-N-acetylglucosamine and FUDP-glucose. No difference was observed in both conditions of culture. The viability of yeast cells on treatment with 5-fluorouracil was also followed. Both FUDP-sugars were partially purified by column chromatography (on Hypersil ODS and Mono Q columns) and characterized by: (i) treatment with alkaline phosphatase (EC 3.1.3.1), snake venom phosphodiesterase (EC 3.1.4.1) and UDP-glucose dehydrogenase (EC 1.1.1.22); (ii) UV spectra; and (iii) matrix-assisted laser desorption/ionization-time of flight mass analysis and 1H-nuclear magnetic resonance spectrometry. The syntheses of both FUDP-sugars were inversely related to the concentration of uracil and directly related to the concentration of 5-fluorouracil in the culture medium. The strain W303-1A, requiring uracil for growth, was useful as a tool to analyze the effect of 5-fluorouracil on nucleotide metabolism.
Günther Sillero
María A
MA
Departamento de Bioquímica, Instituto de Investigaciones Biomédicas Alberto Sols UAM/CSIC, Facultad de Medicina, Madrid, Spain.
Pérez-Zúñiga
Francisco
F
Gomes
Joana
J
de Carvalho
Ana Isabel
AI
Martins
Susana
S
Silles
Eduardo
E
Sillero
Antonio
A
D000332:Aerobiosis; D000469:Alkaline Phosphatase / Q000378:metabolism; D000963:Antimetabolites / Q000494:pharmacology*; D002853:Chromatography, Liquid; D003470:Culture Media / Q000737:chemistry; D005285:Fermentation; D005468:Fluorodeoxyuridylate / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D005472:Fluorouracil / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy; D050296:Microbial Viability; D043264:Phosphodiesterase I / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013057:Spectrum Analysis; D014498:Uracil / Q000032:analysis; D014532:Uridine Diphosphate Glucose / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014533:Uridine Diphosphate Glucose Dehydrogenase / Q000378:metabolism; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D000963:Antimetabolites; D003470:Culture Media; D005468:Fluorodeoxyuridylate; D014537:Uridine Diphosphate N-Acetylglucosamine; D014498:Uracil; C049358:5-fluorouridine diphosphate glucose; D014533:Uridine Diphosphate Glucose Dehydrogenase; D000469:Alkaline Phosphatase; D043264:Phosphodiesterase I; C470790:phosphodiesterase I, snake venom; D005472:Fluorouracil; D014532:Uridine Diphosphate Glucose