The hexokinase 2 protein participates in regulatory DNA-protein complexes necessary for glucose repression of the SUC2 gene in Saccharomyces cerevisiae

被引:81
作者
Herrero, P [1 ]
Martínez-Campa, C [1 ]
Moreno, F [1 ]
机构
[1] Univ Oviedo, Inst Univ Biotecnol Asturias, Dept Bioquim & Biol Mol, E-33006 Oviedo, Spain
关键词
HXK2; SUC2; glucose repression; Saccharomyces cerevisiae;
D O I
10.1016/S0014-5793(98)00872-2
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The HXK2 gene plays an important role in glucose repression in the yeast Saccharomyces cerevisiae. Recently me have described that the HXK2 gene product, isoenzyme 2 of hexokinase, is located both in the nucleus and in the cytoplasm of S, cerevisiae cells. In this work we used deletion analysis to identify the essential part of the protein-mediating nuclear localisation, Determinations of fructose-kinase activity and immunoblot analysis using anti-Hxk2 antibodies in isolated nuclei, together with observations of the fluorescence distribution of Hxk2-GFP fusion protein in cells transformed with an HXK2::gfp mutant gene, indicated that the decapeptide KKPQARKGSM, located between amino acid residues 7 and 16 of hexokinase 2, is important for nuclear localisation of the protein. Further experimental evidence, measuring invertase activity in mild-type and mutant cells expressing a truncated version of the Hxk2 protein unable to enter the nucleus, shove's that a nuclear localisation of Hxk2 is necessary for glucose repression signalling of the SUC2 gene. Furthermore, we demonstrate using gel mobility shift analysis that Hxk2 participates in DNA-protein complexes,vith cis-acting regulatory elements of the SUC2 gene promoter. (C) 1998 Federation of European Biochemical Societies.
引用
收藏
页码:71 / 76
页数:6
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