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Molecular cloning of CIF1 , a yeast gene necessary for growth on glucose
Author(s) -
González M. Isabel,
Blázquez Miguel A.,
Gancedo Carlos,
Stucka Rolf,
Feldmann Horst
Publication year - 1992
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320080304
Subject(s) - biology , cloning (programming) , molecular cloning , yeast , gene , genetics , computational biology , gene expression , computer science , programming language
The cif1 mutation of Saccharomyces cerevisia (Navon et al., Biochemistry 18 , 4487–4499, 1979) causes inability to grow on glucose and absence of catabolite inactivation. We have cloned the CIF1 gene by complementation of funcion and licated it in a 2·75 kb Sph I‐ Bst EII fragment situated at ca. 18 kb centomere distal of LYS2 and ca. 80 kb centromere proximal of TYRI on chromosome II. Southern analysis demostrated that CIF1 is present in a single copy in the yeast genome. Northern analysis revealed that the corresponding mRNA of 1·8 kb is more abundant in cells grown on galactose than in those grown on glucose. A protein of ca. 54 kDa was predicted from the open reading frame in the sequenced fragment. In strains carrying the cif1 mutation the intracellular concentration of ATP decreased immediately after addition of glucose while the intracellular concentration of cAMP did not increse. cAMP concentration increases in response to galactose or 2,4‐dinitrophenol. Disruption of BCY1 or overexpression of CDC25 in a cif1/ , background did not restore growth on glucose, suggesting that the absence of cAMP signal is not primary cause of lack of growth on glucose. Complementation tests showed that cif1 is not allelic to fdp1 although the two genes seem to be functionally related.