Chatfield-Reed Kate, Vachon Lianne, Kwon Eun-Joo Gina, Chua Gordon
Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada.
Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada
Genetics. 2016 Apr;202(4):1365-75. doi: 10.1534/genetics.115.184218. Epub 2016 Feb 19.
Gene regulation in response to intracellular calcium is mediated by the calcineurin-activated transcription factor Prz1 in the fission yeast Schizosaccharomyces pombe Genome-wide studies of the Crz1 and CrzA fungal orthologs have uncovered numerous target genes involved in conserved and species-specific cellular processes. In contrast, very few target genes of Prz1 have been published. This article identifies an extensive list of genes using transcriptome and ChIP-chip analyses under inducing conditions of Prz1, including CaCl2 and tunicamycin treatment, as well as a ∆pmr1 genetic background. We identified 165 upregulated putative target genes of Prz1 in which the majority contained a calcium-dependent response element in their promoters, similar to that of the Saccharomyces cerevisiae ortholog Crz1 These genes were functionally enriched for Crz1-conserved processes such as cell-wall biosynthesis. Overexpression of prz1(+)increased resistance to the cell-wall degradation enzyme zymolyase, likely from upregulation of theO-mannosyltransferase encoding gene omh1(+) Loss of omh1(+)abrogates this phenotype. We uncovered a novel inhibitory role in flocculation for Prz1. Loss of prz1(+)resulted in constitutive flocculation and upregulation of genes encoding the flocculins Gsf2 and Pfl3, as well as the transcription factor Cbf12. The constitutive flocculation of the ∆prz1 strain was abrogated by the loss of gsf2(+) or cbf12(+) This study reveals that Prz1 functions as a positive and negative transcriptional regulator of genes involved in cell-wall biosynthesis and flocculation, respectively. Moreover, comparison of target genes between Crz1/CrzA and Prz1 indicate some conservation in DNA-binding specificity, but also substantial rewiring of the calcineurin-mediated transcriptional regulatory network.
在裂殖酵母粟酒裂殖酵母中,细胞内钙响应的基因调控由钙调神经磷酸酶激活的转录因子Prz1介导。对Crz1和CrzA真菌直系同源物的全基因组研究发现了许多参与保守和物种特异性细胞过程的靶基因。相比之下,Prz1的靶基因很少被报道。本文在Prz1的诱导条件下,包括氯化钙和衣霉素处理以及∆pmr1遗传背景下,使用转录组和芯片分析鉴定了大量基因。我们鉴定了165个上调的Prz1推定靶基因,其中大多数在其启动子中含有钙依赖性反应元件,类似于酿酒酵母直系同源物Crz1。这些基因在功能上富集于Crz1保守过程,如细胞壁生物合成。prz1(+)的过表达增加了对细胞壁降解酶溶菌酶的抗性,这可能是由于编码O-甘露糖基转移酶的基因omh1(+)的上调。omh1(+)的缺失消除了这种表型。我们发现Prz1在絮凝中具有新的抑制作用。prz1(+)的缺失导致组成型絮凝以及编码絮凝蛋白Gsf2和Pfl3以及转录因子Cbf12的基因上调。∆prz1菌株的组成型絮凝被gsf2(+)或cbf12(+)的缺失所消除。这项研究表明,Prz1分别作为参与细胞壁生物合成和絮凝的基因的正转录调节因子和负转录调节因子发挥作用。此外,Crz1/CrzA和PrzZ靶基因的比较表明,在DNA结合特异性上有一些保守性,但钙调神经磷酸酶介导的转录调控网络也有大量的重新布线。