Department of Biological Sciences, Northern Kentucky University, Highland Heights, KY 41099, USA.
G3 (Bethesda). 2024 Apr 3;14(4). doi: 10.1093/g3journal/jkae002.
The SAM1 and SAM2 genes encode for S-Adenosylmethionine (AdoMet) synthetase enzymes, with AdoMet serving as the main cellular methyl donor. We have previously shown that independent deletion of these genes alters chromosome stability and AdoMet concentrations in opposite ways in Saccharomyces cerevisiae. To characterize other changes occurring in these mutants, we grew wildtype, sam1Δ/sam1Δ, and sam2Δ/sam2Δ strains in 15 different Phenotypic Microarray plates with different components and measured growth variations. RNA-Sequencing was also carried out on these strains and differential gene expression determined for each mutant. We explored how the phenotypic growth differences are linked to the altered gene expression, and hypothesize mechanisms by which loss of the SAM genes and subsequent AdoMet level changes, impact pathways and processes. We present 6 stories, discussing changes in sensitivity or resistance to azoles, cisplatin, oxidative stress, arginine biosynthesis perturbations, DNA synthesis inhibitors, and tamoxifen, to demonstrate the power of this novel methodology to broadly profile changes due to gene mutations. The large number of conditions that result in altered growth, as well as the large number of differentially expressed genes with wide-ranging functionality, speaks to the broad array of impacts that altering methyl donor abundance can impart. Our findings demonstrate that some cellular changes are directly related to AdoMet-dependent methyltransferases and AdoMet availability, some are directly linked to the methyl cycle and its role in production of several important cellular components, and others reveal impacts of SAM gene mutations on previously unconnected pathways.
SAM1 和 SAM2 基因编码 S-腺苷甲硫氨酸(AdoMet)合成酶,AdoMet 作为主要的细胞甲基供体。我们之前的研究表明,这些基因的独立缺失以相反的方式改变了酿酒酵母的染色体稳定性和 AdoMet 浓度。为了研究这些突变体中发生的其他变化,我们在 15 种不同的表型微阵列平板中培养野生型、sam1Δ/sam1Δ 和 sam2Δ/sam2Δ 菌株,平板中含有不同的成分,并测量生长变化。我们还对这些菌株进行了 RNA 测序,并确定了每个突变体的差异基因表达。我们探讨了表型生长差异与改变的基因表达之间的联系,并假设了 SAM 基因缺失和随后的 AdoMet 水平变化影响途径和过程的机制。我们提出了 6 个故事,讨论了对唑类药物、顺铂、氧化应激、精氨酸生物合成扰动、DNA 合成抑制剂和他莫昔芬的敏感性或耐药性变化,以展示这种新方法广泛分析基因突变引起的变化的能力。大量导致生长改变的条件,以及数量众多的具有广泛功能的差异表达基因,说明了改变甲基供体丰度可能产生的广泛影响。我们的研究结果表明,一些细胞变化直接与 AdoMet 依赖性甲基转移酶和 AdoMet 的可用性有关,一些与甲基循环及其在几种重要细胞成分生产中的作用直接相关,其他变化则揭示了 SAM 基因突变对以前不相关途径的影响。