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拟南芥伴侣蛋白复合体CAF-1缺失会导致有丝分裂染色体异常以及参与DNA修复的基因表达谱变化。

The Absence of the Arabidopsis Chaperone Complex CAF-1 Produces Mitotic Chromosome Abnormalities and Changes in the Expression Profiles of Genes Involved in DNA Repair.

作者信息

Varas Javier, Santos Juan L, Pradillo Mónica

机构信息

Departamento de Genética, Facultad de Biología, Universidad Complutense de MadridMadrid, Spain.

出版信息

Front Plant Sci. 2017 Apr 11;8:525. doi: 10.3389/fpls.2017.00525. eCollection 2017.

Abstract

Chromatin Assembly Factor 1 (CAF-1) is an evolutionary conserved heterotrimeric chaperone complex that facilitates the incorporation of histones H3 and H4 onto newly synthesized DNA. We demonstrate here that the mutant deficient for the large subunit of the complex, , and in minor extent, the mutant deficient for the middle subunit, , display chromosome abnormalities throughout Arabidopsis mitosis. Among them, we observed multicentromeric chromosomes at metaphase, and chromatid bridges and acentric fragments at anaphase-telophase. 45S rDNA and telomeric sequences were frequently involved in bridges and fragments. Gene expression analysis by real-time qPCR has revealed that several genes related to homologous recombination (HR) and alternative non-homologous end-joining (aNHEJ) are overexpressed in . These results concur with previous studies which have indicated that HR may be involved in the progressive loss of 45S rDNA and telomeres displayed by mutants. However, increased expression of , and in , and the phenotype shown by the double mutant suggest that aNHEJ should also be responsible for the chromosomal aberrations observed. The activity of different DNA repair pathways in absence of CAF-1 is discussed.

摘要

染色质组装因子1(CAF-1)是一种进化上保守的异源三聚体伴侣复合物,它有助于将组蛋白H3和H4整合到新合成的DNA上。我们在此证明,该复合物大亚基缺陷的突变体,以及在较小程度上,中间亚基缺陷的突变体,在拟南芥有丝分裂全过程中均表现出染色体异常。其中,我们在中期观察到多着丝粒染色体,在后期 - 末期观察到染色单体桥和无着丝粒片段。45S核糖体DNA和端粒序列经常参与桥和片段的形成。通过实时定量PCR进行的基因表达分析表明,几个与同源重组(HR)和替代性非同源末端连接(aNHEJ)相关的基因在该突变体中过表达。这些结果与先前的研究一致,先前的研究表明HR可能参与了该突变体所显示的45S核糖体DNA和端粒的逐渐丢失。然而,该突变体中、和的表达增加,以及双突变体所显示的表型表明,aNHEJ也应该对观察到的染色体畸变负责。本文讨论了在缺乏CAF-1的情况下不同DNA修复途径的活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5195/5386969/7541f87e83ca/fpls-08-00525-g001.jpg

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