Brown T A, Evangelista C, Trumpower B L
Dartmouth Medical School, Department of Biochemistry, Hanover, New Hampshire 03755, USA.
J Bacteriol. 1995 Dec;177(23):6836-43. doi: 10.1128/jb.177.23.6836-6843.1995.
Selection for mutants which release glucose repression of the CYB2 gene was used to identify genes which regulate repression of mitochondrial biogenesis. We have identified two of these as the previously described GRR1/CAT80 and ROX3 genes. Mutations in these genes not only release glucose repression of CYB2 but also generally release respiration of the mutants from glucose repression. In addition, both mutants are partially defective in CYB2 expression when grown on nonfermentable carbon sources, indicating a positive regulatory role as well. ROX3 was cloned by complementation of a glucose-inducible flocculating phenotype of an amber mutant and has been mapped as a new leftmost marker on chromosome 2. The ROX3 mutant has only a modest defect in glucose repression of GAL1 but is substantially compromised in galactose induction of GAL1 expression. This mutant also has increased SUC2 expression on nonrepressing carbon sources. We have also characterized the regulation of CYB2 in strains carrying null mutation in two other glucose repression genes, HXK2 and SSN6, and show that HXK2 is a negative regulator of CYB2, whereas SSN6 appears to be a positive effector of CYB2 expression.
通过筛选能解除CYB2基因葡萄糖阻遏作用的突变体来鉴定调控线粒体生物合成阻遏的基因。我们已确定其中两个基因为先前描述的GRR1/CAT80和ROX3基因。这些基因中的突变不仅能解除CYB2的葡萄糖阻遏,还通常能使突变体的呼吸作用从葡萄糖阻遏中释放出来。此外,当在非发酵性碳源上生长时,这两种突变体在CYB2表达上都存在部分缺陷,这也表明它们具有正向调节作用。ROX3是通过对琥珀突变体的葡萄糖诱导絮凝表型进行互补克隆得到的,并已被定位为2号染色体上最左边的一个新标记。ROX3突变体在GAL1的葡萄糖阻遏方面只有适度缺陷,但在GAL1表达的半乳糖诱导方面却严重受损。该突变体在非阻遏性碳源上SUC2表达也增加。我们还对另外两个葡萄糖阻遏基因HXK2和SSN6携带无效突变的菌株中CYB2的调控进行了表征,结果表明HXK2是CYB2的负调控因子,而SSN6似乎是CYB2表达的正向效应因子。