Cloning and mutational analysis of the gene encoding subunit C of yeast vacuolar H(+)-ATPase.

摘要: A DNA fragment containing the gene encoding subunit C of vaculor H(+)-ATPase (V-ATPase) was cloned from a yeast library. The predicted amino acid sequence indicated that consists 373 acids with calculated molecular mass 42,287 Da. protein is 37 identical in its to bovine V-ATPase. this study contained two additional reading frames. At 5' end an homologous Artemia elongation factor 1 detected. 3' N-terminal part kinesin-like observed. V-ATPase interrupted, and resulting mutant could not grow at high pH sensitive low Ca2+ concentrations growth medium. Transformation by plasmid repaired phenotype mutant. Substitution more than half coding region corresponding resulted indistinguishable wild type. Immunological studies disruptant revealed necessary for assembly catalytic sector enzyme.