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Arabidopsis CHROMOSOME TRANSMISSION FIDELITY 7 (AtCTF7/ECO1) is required for DNA repair, mitosis and meiosisand development through alternative promoters

机译:arabidopsis CHROmOsOmE TRaNsmIssION FIDELITY 7 (atCTF7/ECO1) is required for DNa repair, mitosis and meiosisand development through alternative promoters

摘要

The proper transmission of DNA in dividing cells is crucial for the survival of eukaryotic organisms. During cell division, faithful segregation of replicated chromosomes requires their tight attachment, known as sister chromatid cohesion, until anaphase. Sister chromatid cohesion is established during S-phase in a process requiring an acetyltransferase that in yeast is known as Establishment of cohesion 1 (Eco1). Inactivation of Eco1 typically disrupts chromosome segregation and homologous recombination-dependent DNA repair in dividing cells, ultimately resulting in lethality. We report here the isolation and detailed characterization of two homozygous T-DNA insertion mutants for the Arabidopsis thaliana Eco1 homolog, CHROMOSOME TRANSMISSION FIDELITY 7/ESTABLISHMENT OF COHESION 1 (CTF7/ECO1), called ctf7-1 and ctf7-2. Mutants exhibited dwarfism, poor anther development and sterility. Analysis of somatic tissues by flow cytometry, scanning electron microscopy and quantitative real-time PCR identified defects in DNA repair and cell division, including an increase in the area of leaf epidermal cells, an increase in DNA content and the upregulation of genes involved in DNA repair including BRCA1 and PARP2. No significant change was observed in the expression of genes that influence entry into the endocycle. Analysis of meiocytes identified changes in chromosome morphology and defective segregation; the abundance of chromosomal-bound cohesion subunits was also reduced. Transcript levels for several meiotic genes, including the recombinase genes DMC1 andRAD51C and the S-phase licensing factor CDC45 were elevated in mutant anthers. Taken together our results demonstrate that Arabidopsis CTF7/ECO1 plays important roles in the preservation of genome integrity and meiosis.
机译:DNA在分裂细胞中的正确传播对于真核生物的生存至关重要。在细胞分裂过程中,复制染色体的忠实分离要求它们紧密结合,称为姐妹染色单体凝聚,直到后期。姐妹染色单体内聚力是在S阶段建立的,该过程需要乙酰转移酶,而在酵母中这种酶被称为内聚力建立1(Eco1)。 Eco1的失活通常会破坏分裂细胞中的染色体分离和依赖同源重组的DNA修复,最终导致致死性。我们在此报告了两个拟南芥Eco1同源纯合T-DNA插入突变体的色谱分离和详细表征,染色体传递保真度7 /建立凝聚力1(CTF7 / ECO1),分别称为ctf7-1和ctf7-2。突变体表现出矮化,花药发育不良和不育。通过流式细胞仪,扫描电子显微镜和定量实时PCR对体组织进行分析,发现了DNA修复和细胞分裂的缺陷,包括叶表皮细胞面积的增加,DNA含量的增加以及与DNA有关的基因的上调维修包括BRCA1和PARP2。没有观察到影响进入内循环的基因表达的显着变化。通过对单核细胞的分析,可以确定染色体形态的变化和缺陷性的分离;染色体结合内聚亚基的丰度也降低了。突变体花药中包括减数分裂基因包括重组酶基因DMC1和RAD51C以及S期许可因子CDC45的转录水平升高。综合我们的结果表明,拟南芥CTF7 / ECO1在保存基因组完整性和减数分裂中起重要作用。

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