Schüller H J, Entian K D
Institute for Microbiology, Johann Wolfgang Goethe-Universität, Frankfurt am Main, Federal Republic of Germany.
J Bacteriol. 1991 Mar;173(6):2045-52. doi: 10.1128/jb.173.6.2045-2052.1991.
Saccharomyces cerevisiae regulatory genes CAT1 and CAT3 constitute a positive control circuit necessary for derepression of gluconeogenic and disaccharide-utilizing enzymes. Mutations within these genes are epistatic to hxk2 and hex2, which cause defects in glucose repression. cat1 and cat3 mutants are unable to grow in the presence of nonfermentable carbon sources or maltose. Stable gene disruptions were constructed inside these genes, and the resulting growth deficiencies were used for selecting epistatic mutations. The revertants obtained were tested for glucose repression, and those showing altered regulatory properties were further investigated. Most revertants belonged to a single complementation group called cat4. This recessive mutation caused a defect in glucose repression of invertase, maltase, and iso-1-cytochrome c. Additionally, hexokinase activity was increased. Gluconeogenic enzymes are still normally repressible in cat4 mutants. The occurrence of recombination of cat1::HIS3 and cat3::LEU2 with some cat4 alleles allowed significant growth in the presence of ethanol, which could be attributed to a partial derepression of gluconeogenic enzymes. The cat4 complementation group was tested for allelism with hxk2, hex2, cat80, cid1, cyc8, and tup1 mutations, which were previously described as affecting glucose repression. Allelism tests and tetrad analysis clearly proved that the cat4 complementation group is a new class of mutant alleles affecting carbon source-dependent gene expression.
酿酒酵母调控基因CAT1和CAT3构成了一个正调控回路,该回路对于解除糖异生酶和双糖利用酶的阻遏是必需的。这些基因内的突变对hxk2和hex2呈上位性,hxk2和hex2会导致葡萄糖阻遏缺陷。cat1和cat3突变体在非发酵性碳源或麦芽糖存在的情况下无法生长。在这些基因内部构建了稳定的基因破坏,并且利用由此产生的生长缺陷来选择上位性突变。对获得的回复突变体进行葡萄糖阻遏测试,并对那些显示出调控特性改变的回复突变体进行进一步研究。大多数回复突变体属于一个称为cat4的单一互补群。这种隐性突变导致蔗糖酶、麦芽糖酶和同工-1-细胞色素c的葡萄糖阻遏存在缺陷。此外,己糖激酶活性增加。在cat4突变体中,糖异生酶仍然可以正常被阻遏。cat1::HIS3和cat3::LEU2与一些cat4等位基因发生重组,使得在乙醇存在的情况下能够显著生长,这可能归因于糖异生酶的部分去阻遏。对cat4互补群进行了与hxk2、hex2、cat80、cid1、cyc8和tup1突变的等位性测试,这些突变先前被描述为影响葡萄糖阻遏。等位性测试和四分体分析清楚地证明,cat4互补群是一类影响碳源依赖性基因表达的新的突变等位基因。