Expression of human RECQL5 in Saccharomyces cerevisiae causes transcription defects and transcription-associated genome instability.

IF 2.3 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Juan Lafuente-Barquero, Jesper Q Svejstrup, Rosa Luna, Andrés Aguilera
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Abstract

RECQL5 is a member of the conserved RecQ family of DNA helicases involved in the maintenance of genome stability that is specifically found in higher eukaryotes and associates with the elongating RNA polymerase II. To expand our understanding of its function we expressed human RECQL5 in the yeast Saccharomyces cerevisiae, which does not have a RECQL5 ortholog. We found that RECQL5 expression leads to cell growth inhibition, increased genotoxic sensitivity and transcription-associated hyperrecombination. Chromatin immunoprecipitation and transcriptomic analysis of yeast cells expressing human RECQL5 shows that this is recruited to transcribed genes and although it causes only a weak impact on gene expression, in particular at G + C-rich genes, it leads to a transcription termination defect detected as readthrough transcription. The data indicate that the interaction between RNAPII and RECQL5 is conserved from yeast to humans. Unexpectedly, however, the RECQL5-ID mutant, previously shown to have reduced the association with RNAPII in vitro, associates with the transcribing polymerase in cells. As a result, expression of RECQL5-ID leads to similar although weaker phenotypes than wild-type RECQL5 that could be transcription-mediated. Altogether, the data suggests that RECQL5 has the intrinsic ability to function in transcription-dependent and independent genome dynamics in S. cerevisiae.

Abstract Image

在酿酒酵母中表达人类 RECQL5 会导致转录缺陷和与转录相关的基因组不稳定性。
RECQL5是DNA螺旋酶保守RecQ家族的一个成员,它参与维持基因组的稳定性,专门存在于高等真核生物中,并与延伸RNA聚合酶II相关联。为了扩大对其功能的了解,我们在没有 RECQL5 同源物的酵母中表达了人类 RECQL5。我们发现,RECQL5 的表达会导致细胞生长抑制、基因毒性敏感性增加以及转录相关的过度重组。对表达人 RECQL5 的酵母细胞进行的染色质免疫沉淀和转录组分析表明,RECQL5 被转录到转录基因上,虽然它对基因表达的影响很弱,特别是在富含 G + C 的基因上,但它会导致转录终止缺陷,被检测为穿透转录。这些数据表明,从酵母到人类,RNAPII 和 RECQL5 之间的相互作用是保守的。然而,令人意想不到的是,以前在体外与 RNAPII 的结合减少的 RECQL5-ID 突变体,在细胞中也与转录聚合酶结合。因此,与野生型 RECQL5 相比,RECQL5-ID 的表达会导致类似但较弱的表型,这些表型可能是转录介导的。总之,这些数据表明,RECQL5 在 S. cerevisiae 中具有依赖转录和独立基因组动态的内在能力。
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来源期刊
Molecular Genetics and Genomics
Molecular Genetics and Genomics 生物-生化与分子生物学
CiteScore
5.10
自引率
3.20%
发文量
134
审稿时长
1 months
期刊介绍: Molecular Genetics and Genomics (MGG) publishes peer-reviewed articles covering all areas of genetics and genomics. Any approach to the study of genes and genomes is considered, be it experimental, theoretical or synthetic. MGG publishes research on all organisms that is of broad interest to those working in the fields of genetics, genomics, biology, medicine and biotechnology. The journal investigates a broad range of topics, including these from recent issues: mechanisms for extending longevity in a variety of organisms; screening of yeast metal homeostasis genes involved in mitochondrial functions; molecular mapping of cultivar-specific avirulence genes in the rice blast fungus and more.
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