组蛋白变异H3.5在睾丸细胞分化中的作用及其与组蛋白伴侣的相互作用。

IF 3.8 2区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Patrick Philipp Weil, Anton Pembaur, Beatrice Wirth, Eda Oetjen, Hannes Büsscher, Klemens Zirngibl, Malte Czarnetzki, Stella Braun, Jann-Frederik Cremers, Daniel Gödde, Stephan Degener, Jan Postberg
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引用次数: 0

摘要

睾丸细胞分化是一个高度调控的过程,对男性生殖健康至关重要。组蛋白变体H3.5显然在这种复杂的细胞类型管弦乐队中起着关键作用,但其调控和功能仍然知之甚少。为了阐明其作用,我们使用c-Kit/CD117作为分离标记物对睾丸细胞进行分离并分析H3.5的表达。此外,我们利用公共数据库研究了H3.5表达的调控。我们探索了H3-5基因特定区域的DNA甲基化模式,并评估了精原细胞瘤标本中H3-5拷贝数的增加。此外,我们检测了H3.5的睾丸定位及其组蛋白伴侣的相互作用,以了解其在蛋白质水平上的调控。我们采用qRT-PCR、MeDIP和qPCR技术研究了H3.5在不同细胞类型中的表达和DNA甲基化。采用qPCR分析H3-5拷贝数增益。用荧光-2杂交法研究了幼鼠肾细胞中蛋白质的相互作用。H3.5主要富集于精母细胞。与H3-5启动子重叠的CpG岛的DNA甲基化似乎参与了H3.5表达的组织特异性调控。在精原细胞瘤标本中观察到H3.5表达升高,提示其与睾丸肿瘤的潜在联系。H3-5拷贝数增加与精原细胞瘤标本中H3.5表达升高相关。此外,我们确定了H3.5与组蛋白伴侣Asf1a和Asf1b、HIRA、CAF p150和DAXX之间的物理相互作用,揭示了H3.5的蛋白水平调控。这些发现为睾丸细胞分化的分子机制和H3.5在睾丸病理中的潜在作用提供了有价值的见解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones.

Testicular cell differentiation is a highly regulated process, essential for male reproductive health. The histone variant H3.5 is apparently a critical player in this intricate orchestra of cell types, but its regulation and function remains poorly understood. To elucidate its role, we fractionized testicular cells using c-Kit/CD117 as a separation marker and analyzed H3.5 expression. Further, we investigated the regulation of H3.5 expression using public data repositories. We explored DNA methylation patterns in specific regions of the H3-5 gene and assessed H3-5 copy number gain in seminoma specimens. Additionally, we examined the testicular localization of H3.5 and its histone chaperone interactions to understand its regulation at the protein level. We used qRT-PCR, MeDIP, and qPCR to study H3.5 expression and DNA methylation in various cell types. H3-5 copy number gain was analyzed using qPCR. Protein interactions were investigated through fluorescence-2-hybrid assays in baby hamster kidney cells. H3.5 is primarily enriched in spermatocytes. DNA methylation of a CpG island overlapping the H3-5 promoter appeared to be involved in the tissue-specific regulation of H3.5 expression. Elevated H3.5 expression was observed in seminoma specimens, suggesting a potential link to testicular tumors. H3-5 copy number gain was associated with elevated H3.5 expression in seminoma specimens. Furthermore, we identified physical interactions between H3.5 and histone chaperones Asf1a and Asf1b, HIRA, CAF p150 and DAXX, shedding light on the protein-level regulation of H3.5. These findings provide valuable insights into the molecular mechanisms governing testicular cell differentiation and the potential role of H3.5 in testicular pathologies.

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来源期刊
Scientific Reports
Scientific Reports Natural Science Disciplines-
CiteScore
7.50
自引率
4.30%
发文量
19567
审稿时长
3.9 months
期刊介绍: We publish original research from all areas of the natural sciences, psychology, medicine and engineering. You can learn more about what we publish by browsing our specific scientific subject areas below or explore Scientific Reports by browsing all articles and collections. Scientific Reports has a 2-year impact factor: 4.380 (2021), and is the 6th most-cited journal in the world, with more than 540,000 citations in 2020 (Clarivate Analytics, 2021). •Engineering Engineering covers all aspects of engineering, technology, and applied science. It plays a crucial role in the development of technologies to address some of the world''s biggest challenges, helping to save lives and improve the way we live. •Physical sciences Physical sciences are those academic disciplines that aim to uncover the underlying laws of nature — often written in the language of mathematics. It is a collective term for areas of study including astronomy, chemistry, materials science and physics. •Earth and environmental sciences Earth and environmental sciences cover all aspects of Earth and planetary science and broadly encompass solid Earth processes, surface and atmospheric dynamics, Earth system history, climate and climate change, marine and freshwater systems, and ecology. It also considers the interactions between humans and these systems. •Biological sciences Biological sciences encompass all the divisions of natural sciences examining various aspects of vital processes. The concept includes anatomy, physiology, cell biology, biochemistry and biophysics, and covers all organisms from microorganisms, animals to plants. •Health sciences The health sciences study health, disease and healthcare. This field of study aims to develop knowledge, interventions and technology for use in healthcare to improve the treatment of patients.
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