Linden H, Macino G
Dipartimento di Biopatologia Umana, Sezione Biologia Cellulare, Università di Roma La Sapienza, Italy.
EMBO J. 1997 Jan 2;16(1):98-109. doi: 10.1093/emboj/16.1.98.
A saturating genetic dissection of 'blind' mutants in Neurospora crassa has identified a total of two non-redundant loci (wc-1 and wc-2) each of which is required for blue-light perception/signal transduction. Previously, we demonstrated that WC1 is a putative zinc finger transcription factor able to bind specifically to a light-regulated promoter. Here, we present the cloning and characterization of the wc-2 gene. We demonstrate using mutation analysis and in vitro DNA-binding assays that WC2, the second partner of this light signal transduction system, encodes a functional zinc finger DNA-binding protein with putative PAS dimerization and transcription activation domains. This molecular genetic dissection of the second of two components of this light signal transduction system has enabled us to devise a model whereby WC1 and WC2 are proposed to interact via homologous PAS domains, bind to promoters of light-regulated genes and activate transcription. As such, this study provides the first insight into two co-operating partners in blue-light signal transduction in any organism and describes the molecular tools with which to dissect this enigmatic process.
对粗糙脉孢菌“盲目”突变体进行的饱和遗传剖析总共鉴定出两个非冗余基因座(wc-1和wc-2),蓝光感知/信号转导过程中每个基因座都是必需的。此前,我们证明WC1是一种推定的锌指转录因子,能够特异性结合光调节启动子。在此,我们展示了wc-2基因的克隆和特性分析。我们通过突变分析和体外DNA结合试验证明,该光信号转导系统的第二个组分WC2编码一种功能性锌指DNA结合蛋白,具有推定的PAS二聚化和转录激活结构域。对该光信号转导系统两个组分中第二个组分的这种分子遗传学剖析,使我们能够设计出一个模型,据此提出WC1和WC2通过同源PAS结构域相互作用,结合光调节基因的启动子并激活转录。因此,本研究首次深入了解了任何生物体蓝光信号转导中的两个协同作用伙伴,并描述了剖析这一神秘过程的分子工具。