Remines McKayla, Schoonover Makailyn, Knox Zoey, Kenwright Kailee, Hoffert Kellyn M, Coric Amila, Mead James, Ampfer Joseph, Seye Serigne, Strome Erin D
Department of Biological Sciences, Northern Kentucky University, Highland Heights, KY 41099.
bioRxiv. 2023 Jun 10:2023.06.09.544294. doi: 10.1101/2023.06.09.544294.
The and genes encode for S-AdenosylMethionine (AdoMet) synthetase enzymes, with AdoMet serving as the main methyl donor. We have previously shown that independent deletion of these genes alters chromosome stability and AdoMet concentrations in opposite ways in To characterize other changes occurring in these mutants, we grew wildtype, and strains in 15 different Phenotypic Microarray plates with different components, equal to 1440 wells, and measured for growth variations. RNA-Sequencing was also carried out on these strains and differential gene expression determined for each mutant. In this study, we explore how the phenotypic growth differences are linked to the altered gene expression, and thereby predict the mechanisms by which loss of the genes and subsequent AdoMet level changes, impact pathways and processes. We present six 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, even when the conditions tested were not specifically selected as targeting known methyl involving pathways. 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 is production of several important cellular components, and others reveal impacts of gene mutations on previously unconnected pathways.
和基因编码S-腺苷甲硫氨酸（AdoMet）合成酶，AdoMet作为主要的甲基供体。我们之前已经表明，在中独立缺失这些基因会以相反的方式改变染色体稳定性和AdoMet浓度。为了表征这些突变体中发生的其他变化，我们在15种不同成分的表型微阵列平板（共1440个孔）中培养野生型、和菌株，并测量生长变化。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。在本研究中，我们探讨了表型生长差异如何与改变的基因表达相关联，从而预测基因缺失和随后的AdoMet水平变化影响途径和过程的机制。我们讲述了六个事例，讨论了对唑类、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抗性变化，以证明这种新方法在广泛描述基因突变导致的变化方面的能力。导致生长改变的大量条件，以及功能广泛的大量差异表达基因，表明即使所测试的条件并非专门选择针对已知的涉及甲基的途径，改变甲基供体丰度也能产生广泛的影响。我们的研究结果表明，一些细胞变化与依赖AdoMet的甲基转移酶和AdoMet可用性直接相关，一些与甲基循环及其在几种重要细胞成分产生中的作用直接相关，还有一些揭示了基因突变对以前未关联途径的影响。
