Molecular cloning of Drosophila γ‐glutamylcysteine synthetase by functional complementation of a yeast mutant

FEBS Letters

McLellan, Lesley I.¹  Saunders, Robert D.C.² 

[1] Biomedical Research Centre, Ninewells Hospital and Medical School, University of Dundee, Dundee DD1 9SY, UK;Department of Anatomy and Physiology, University of Dundee, Dundee DD1 4HN, UK

关键词: Glutathione;    γ-Glutamylcysteine synthetase;    Functional complementation;    Drosophila melanogaster;   

DOI  :  10.1016/S0014-5793(00)01148-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

γ-Glutamylcysteine synthetase (GCS) catalyses a critical, rate-limiting step in glutathione synthesis. In this study we describe the isolation and characterisation of a GCS cDNA (pDmGCS4.3.3) from Drosophila melanogaster by functional complementation of a Saccharomyces cerevisiae gsh1 mutant. Expression of pDmGCS4.3.3 in the yeast mutant partially restored glutathione levels and conferred resistance to methylglyoxal. The pDmGCS4.3.3 cDNA was found to be approx. 4.6 kb in length, containing a 2 kb fragment encoding an open reading frame with a high degree of deduced amino acid sequence identity with previously reported GCS sequences. In situ hybridisation revealed that the Drosophila GCS gene maps to 7D6–9 on the X chromosome.

【 授权许可】

Unknown