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编码酵母锌簇蛋白的基因的表型分析。

Phenotypic analysis of genes encoding yeast zinc cluster proteins.

作者信息

Akache B, Wu K, Turcotte B

机构信息

Department of Medicine, Royal Victoria Hospital, McGill University, 687 Pine Avenue West, Montréal, Québec H3A 1A1, Canada.

出版信息

Nucleic Acids Res. 2001 May 15;29(10):2181-90. doi: 10.1093/nar/29.10.2181.

Abstract

Zinc cluster proteins (or binuclear cluster proteins) possess zinc fingers of the Zn(II)2Cys6-type involved in DNA recognition as exemplified by the well-characterized protein Gal4p. These fungal proteins are transcriptional regulators of genes involved in a wide variety of cellular processes including metabolism of compounds such as amino acids and sugars, as well as control of meiosis, multi-drug resistance etc. The yeast (Saccharomyces cerevisiae) sequencing project has allowed the identification of additional zinc cluster proteins for a total of 54. However, the role of many of these putative zinc cluster proteins is unknown. We have performed phenotypic analysis of 33 genes encoding (putative) zinc cluster proteins. Only two members of the GAL4 family are essential genes. Our results show that deletion of eight different zinc cluster genes impairs growth on non-fermentable carbon sources. The same strains are also hypersensitive to the antifungal calcofluor white suggesting a role for these genes in cell wall integrity. In addition, one of these strains (YFL052W) is also heat sensitive on rich (but not minimal) plates. Thus, deletion of YFL052W results in sensitivity to a combination of low osmolarity and high temperature. In addition, six strains are hypersensitive to caffeine, an inhibitor of the MAP kinase pathway and phosphodiesterase of the cAMP pathway. In conclusion, our analysis assigns phenotypes to a number of genes and provides a basis to better understand the role of these transcriptional regulators.

摘要

锌簇蛋白(或双核簇蛋白)具有参与DNA识别的Zn(II)2Cys6型锌指结构,典型代表为已被充分研究的Gal4p蛋白。这些真菌蛋白是参与多种细胞过程的基因的转录调节因子,这些过程包括氨基酸和糖类等化合物的代谢,以及减数分裂控制、多药耐药性等。酵母(酿酒酵母)测序项目已鉴定出另外54种锌簇蛋白。然而,这些推定的锌簇蛋白中许多的作用尚不清楚。我们对33个编码(推定的)锌簇蛋白的基因进行了表型分析。GAL4家族中只有两个成员是必需基因。我们的结果表明,八个不同锌簇基因的缺失会损害在非发酵碳源上的生长。相同的菌株对抗真菌剂荧光增白剂也高度敏感,表明这些基因在细胞壁完整性方面发挥作用。此外,其中一个菌株(YFL052W)在丰富(但不是基本)培养基平板上也对热敏感。因此,YFL052W的缺失导致对低渗透压和高温组合敏感。此外,六个菌株对咖啡因高度敏感,咖啡因是MAP激酶途径和cAMP途径磷酸二酯酶的抑制剂。总之,我们的分析为许多基因赋予了表型,并为更好地理解这些转录调节因子的作用提供了基础。

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