College of Life Sciences, Shanghai Normal University, Shanghai, 200234, China; Department of Obstetrics and Gynecology, Shanghai Everjoy Medical Polyclinic, 675 Minbei Road, Shanghai, 201107, China.
College of Life Sciences, Shanghai Normal University, Shanghai, 200234, China.
J Trace Elem Med Biol. 2021 Jul;66:126759. doi: 10.1016/j.jtemb.2021.126759. Epub 2021 Apr 14.
Acquisition and distribution of zinc supports a number of biological processes. Various molecular factors are involved in zinc metabolism but not fully explored.
Spontaneous mutants were generated in yeast with excess zinc culture followed by whole genome DNA sequencing to discover zinc metabolism related genes by bioinformatics. An identified mutant was characterized through metallomic and molecular biology methods.
Here we reported that MTM1 knockout cells displayed much stronger zinc tolerance than wild type cells on SC medium when exposed to excess zinc. Zn accumulation of mtm1Δ cells was dramatically decreased compared to wild type cells under excessive zinc condition due to MTM1 deletion reduced zinc uptake. ZRC1 mRNA level of mtm1Δ cells was significantly higher than that in the wild-type strain leading to increased vacuolar zinc accumulations in mtm1Δ cells. The mRNA levels of ZRT1 and ZAP1 decreased in mtm1Δ cells contributing to less Zn uptake. The zrc1Δmtm1Δ double knockout strain exhibited Zn sensitivity. MTM1 knockout did not afford resistance to excess zinc through an effect mediated through an influence on levels of ROS. Superoxide dismutase 2 (Sod2p) activity in mtm1Δ cells was severely impaired and not restored through Zn supplementation. Meanwhile, additional Zn showed no significant effect on the localization and expression of Mtm1p.
Our study reveals the MTM1 gene plays an important role in the regulation of zinc homeostasis in yeast cells via changing zinc uptake and distribution. This discovery provides new insights for better understanding biochemical communication between vacuole and mitochondrial in relation to zinc-metabolism.
锌的获取和分布支持许多生物过程。各种分子因素都参与了锌代谢,但尚未完全探索。
在富含锌的培养基中培养酵母,生成自发突变体,然后通过全基因组 DNA 测序,通过生物信息学发现与锌代谢相关的基因。通过金属组学和分子生物学方法对鉴定出的突变体进行表征。
在这里,我们报告说,当暴露于过量锌时,mtm1Δ 细胞比野生型细胞在 SC 培养基上显示出更强的锌耐受性。由于 MTM1 缺失减少了锌的摄取,mtm1Δ 细胞在过量锌条件下的 Zn 积累明显减少。与野生型菌株相比,mtm1Δ 细胞的 ZRC1 mRNA 水平显著升高,导致 mtm1Δ 细胞中液泡锌积累增加。zrt1Δ 和 zap1Δ 细胞中的 ZAP1 mRNA 水平降低,导致 Zn 摄取减少。zrc1Δmtm1Δ 双敲除菌株表现出对 Zn 的敏感性。mtm1Δ 敲除不通过影响 ROS 水平来提供对过量锌的抗性。mtm1Δ 细胞中的超氧化物歧化酶 2(Sod2p)活性严重受损,并且不能通过 Zn 补充来恢复。同时,额外的 Zn 对 Mtm1p 的定位和表达没有显著影响。
我们的研究表明,MTM1 基因通过改变锌的摄取和分布,在酵母细胞中调节锌稳态中发挥重要作用。这一发现为更好地理解与锌代谢相关的液泡和线粒体之间的生化通讯提供了新的见解。