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Utilization of adenine and guanine as nitrogen sources by Chlamydomonas reinhardtii cells
Authors:T A LISA  P PIEDRAS  J CÁRDENAS  M PINEDA
Institution:Departamento de Biaquimica y Bialogi'a Molecular. Facultad de Ciencias, Universidad de Córdoba, 14071-Córdoba, Spain
Abstract:Chlamydomonas reinhardtü Dangeard, adenine or guanine can be used as the sole nitrogen source for growth by means of an inducible system which is repressed by ammonia. Cells grown on either adenine or guanine were able to take up both purines, although the adenine uptake rate was always about 40% of the guanine uptake rate. Both adenine and guanine were taken up by an inducible system(s) exhibiting hyperbolic kinetics with identical apparent A, values of 3-2 mmol m?3 for adenine and 3-2mmol m?3 for guanine. Adenine and guanine utilization depended on pH, with similar optimal pH values of 7·3 and 7·4, respectively. Adenine and guanine each acted as a competitive inhibitor of the other's uptake, and their utilization was also inhibited by hypoxanthine, xanthine and urate. Inhibition of adenine uptake by guanine and hypoxanthine was competitive, with A′, values of 5·5 and 1. 6 mmol m?3 respectively. Guanine uptake was also inhibited competitively by adenine (K1= 1·3mmol m?3) and hypoxanthine (K1= 3. 3 mmol m?3). Utilization of both adenine and guanine was inhibited by cyanide, azide, 3-(3,4-dichlorophenyl)-1,1-dimethyl urea, 2,4-dinitrophenol and carbonylcyanide m-chlorophenylhydrazone, and was also sensitive to p-hydroxymercuribenzoate and N-ethyl-maleimide. On the basis of these results, taken together, the possibility that adenine and guanine are translocated into Chlamydomonas by a common system is discussed.
Keywords:Chlamydomonas            adenine  guanine  nitrogen sources  purine metabolism  purine uptake
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