Hikkel Imrich, Lucau-Danila Ancuta, Delaveau Thierry, Marc Philippe, Devaux Frédéric, Jacq Claude
Laboratoire de Génétique Moléculaire, CNRS UMR8541, Ecole Normale Supérieure, 46 rue d'Ulm 75230 Paris Cedex 05, France.
J Biol Chem. 2003 Mar 28;278(13):11427-32. doi: 10.1074/jbc.M208549200. Epub 2003 Jan 14.
We demonstrate a genomewide approach to determine the physiological role of a putative transcription factor, Ylr266, identified through yeast genome sequencing program. We constructed activated forms of the zinc finger (Zn(2)Cys(6)) protein Ylr266, and we analyzed the corresponding transcriptomes with DNA microarrays to characterize the up-regulated genes. The direct target genes of Ylr266 were further identified by in vivo chromatin immunoprecipitation procedure. The functions of the genes directly controlled by YLR266c are in agreement with the observed drug-resistance phenotype of the cell expressing an activated form of Ylr266. These target genes code for ATP-binding cassette or major facilitator superfamily transporters such as PDR15, YOR1, or AZR1 or for other proteins such as SNG1, YJL216c, or YLL056c which are already known to be involved in the yeast pleiotropic drug resistance (PDR) phenomenon. YLR266c could thus be named PDR8. Overlaps with the other PDR networks argue in favor of a new specific role for PDR8 in connection with the well known PDR regulators PDR1/PDR3 and YRR1. This strategy to identify the regulatory properties of an anonymous transcription factor is likely to be generalized to all the Zn(2)Cys(6) transcription factors from Saccharomyces cerevisiae and related yeasts.
我们展示了一种全基因组方法，以确定通过酵母基因组测序计划鉴定出的假定转录因子Ylr266的生理作用。我们构建了锌指（Zn(2)Cys(6)）蛋白Ylr266的激活形式，并使用DNA微阵列分析相应的转录组，以表征上调基因。通过体内染色质免疫沉淀程序进一步鉴定了Ylr266的直接靶基因。由YLR266c直接控制的基因的功能与表达Ylr266激活形式的细胞所观察到的耐药表型一致。这些靶基因编码ATP结合盒或主要易化子超家族转运蛋白，如PDR15、YOR1或AZR1，或编码其他蛋白质，如SNG1、YJL216c或YLL056c，这些蛋白质已知参与酵母多药耐药（PDR）现象。因此，YLR266c可命名为PDR8。与其他PDR网络的重叠表明PDR8在与众所周知的PDR调节因子PDR1/PDR3和YRR1相关方面具有新的特定作用。这种鉴定无名转录因子调控特性的策略可能会推广到酿酒酵母和相关酵母的所有Zn(2)Cys(6)转录因子。
