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细胞体积增大突变体的表型分析和分子特征研究。

Phenotypic Analysis and Molecular Characterization of Enlarged Cell Size Mutant in .

机构信息

State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361000, China.

College of Ocean and Earth Sciences, Xiamen University, Xiamen 361000, China.

出版信息

Int J Mol Sci. 2023 Sep 2;24(17):13595. doi: 10.3390/ijms241713595.

Abstract

The cell cycle is the fundamental cellular process of eukaryotes. Although cell-cycle-related genes have been identified in microalgae, their cell cycle progression differs from species to species. Cell enlargement in microalgae is an essential biological trait. At the same time, there are various causes of cell enlargement, such as environmental factors, especially gene mutations. In this study, we first determined the phenotypic and biochemical characteristics of a previously obtained enlarged-cell-size mutant of , which was designated . Whole-genome sequencing analysis of the insertion sites of indicated that the insertion fragment is integrated inside the 5'-UTR of U/P-type cyclin and significantly decreases the gene expression of this cyclin. In addition, the transcriptome showed that is a highly expressed cyclin. Furthermore, cell cycle analysis and RT-qPCR of cell-cycle-related genes showed that maintains a high proportion of 4C cells and a low proportion of 1C cells, and the expression level of in wild-type (WT) cells is significantly increased at the end of the light phase and the beginning of the dark phase. This means that is involved in cell division in the dark phase. Our results explain the reason for the larger size. Mutation of leads to the failure of to fully complete cell division in the dark phase, resulting in an enlargement of the cell size and a decrease in cell density, which is helpful to understand the function of in the cell cycle.

摘要

细胞周期是真核生物的基本细胞过程。虽然在微藻中已经鉴定出与细胞周期相关的基因,但它们的细胞周期进程因物种而异。微藻的细胞增大是一种重要的生物学特征。同时,细胞增大有多种原因,如环境因素,特别是基因突变。在本研究中,我们首先确定了先前获得的 细胞大小增大突变体的表型和生化特征,将其命名为 。对 插入位点的全基因组测序分析表明,插入片段整合在 U/P 型细胞周期蛋白 的 5'UTR 内,显著降低了该细胞周期蛋白的基因表达。此外,转录组显示 是一个高度表达的细胞周期蛋白。此外,细胞周期分析和细胞周期相关基因的 RT-qPCR 显示, 在 4C 细胞中保持高比例,在 1C 细胞中保持低比例,并且在光相结束和暗相开始时野生型(WT)细胞中 的表达水平显著增加。这意味着 参与暗相的细胞分裂。我们的结果解释了 尺寸较大的原因。 的突变导致 不能在暗相中完全完成细胞分裂,导致细胞大小增大和细胞密度降低,这有助于理解 在 细胞周期中的功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d6d0/10487731/bd2f6d7fa3bd/ijms-24-13595-g001.jpg

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