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RNA synthesis in liposomes with negatively charged lipids after fusion via freezing-thawing. 带负电荷的脂质体通过冷冻-解冻融合后的 RNA 合成。
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-26 Epub Date: 2024-02-21 DOI: 10.1266/ggs.23-00297
Gakushi Tsuji, Ayu Shimomura, Shota Fukuoka, Masaya Oki

The freezing-thawing (F/T) method for fusing giant unilamellar vesicles (GUVs) can provide substrates, enzymes and membrane material simultaneously and repetitively, and is useful for constructing a recursive model of an artificial cell. However, enzymatic efficiency after F/T is reduced due to rupture of the GUVs and leakage of the inner solution during F/T. Previously, liposomes composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and a negatively charged lipid, such as 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (POPG), showed lower rupture and leakage rates during F/T. In this study, we investigated the effect of POPG on the supply of components required for T7 RNA polymerase reactions via F/T by flow cytometry analysis. We found that the addition of POPG to liposome preparations reduced the efficiency of RNA synthesis. In addition, DNA was concentrated during F/T and RNA synthesis occurred after F/T in liposomes composed of POPC and POPG. Our results provide new insights for more efficient supply of substrates and enzymes by the F/T method, thereby increasing the utility of the F/T method for the construction of recursive bioreactors.

用冷冻-解冻(F/T)方法融合巨型单拉美米尔囊泡(GUVs)可同时重复提供底物、酶和膜材料,有助于构建人工细胞递归模型。然而,在 F/T 过程中,由于 GUV 破裂和内部溶液泄漏,F/T 后的酶效率会降低。以前,由 1-棕榈酰-2-油酰-sn-甘油-3-磷酸胆碱(POPC)和带负电荷的脂质(如 1-棕榈酰-2-油酰-sn-甘油-3-磷酸-(1'-rac-甘油)(POPG)组成的脂质体在 F/T 期间显示出较低的破裂率和泄漏率。在本研究中,我们通过流式细胞仪分析,研究了 POPG 对 T7 RNA 聚合酶反应所需成分供应的影响。我们发现,在脂质体制备过程中加入 POPG 会降低 RNA 合成的效率。此外,在由 POPC 和 POPG 组成的脂质体中,DNA 在 F/T 期间被浓缩,而 RNA 的合成则在 F/T 之后进行。我们的研究结果为利用 F/T 法更有效地提供底物和酶提供了新的启示,从而提高了 F/T 法在构建递归生物反应器方面的实用性。
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引用次数: 0
IMD2, located near the boundary of heterochromatin regions, is regulated by multiple HAT-related factors. IMD2 位于异染色质区域的边界附近,受多种 HAT 相关因子的调控。
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-26 Epub Date: 2024-02-21 DOI: 10.1266/ggs.23-00284
Takahito Ayano, Masaya Oki

In Saccharomyces cerevisiae, boundaries formed by DNA sequence-dependent or -independent histone modifications stop the spread of the heterochromatin region formed via the Sir complex. However, it is unclear whether the histone modifiers that control DNA sequence-independent boundaries function in a chromosome-specific or -nonspecific manner. In this study, we evaluated the effects of the SAGA complex, a histone acetyltransferase (HAT) complex, and its relationship with other histone-modifying enzymes to clarify the mechanism underlying boundary regulation of the IMD2 gene on the right subtelomere of chromosome VIII. We found that Spt8, a component of the SAGA complex, is important for boundary formation in this region and that the inclusion of Spt8 in the SAGA complex is more important than its interaction with TATA-binding protein and TFIIS. In addition to SAGA, various HAT-related factors, such as NuA4 and Rtt109, also functioned in this region. In particular, the SAGA complex induced weak IMD2 expression throughout the cell, whereas NuA4 induced strong expression. These results indicate that multiple HATs contribute to the regulation of boundary formation and IMD2 expression on the right subtelomere of chromosome VIII and that IMD2 expression is determined by the balance between these factors.

在酿酒酵母(Saccharomyces cerevisiae)中,由依赖 DNA 序列或不依赖 DNA 序列的组蛋白修饰形成的边界阻止了通过 Sir 复合物形成的异染色质区域的扩散。然而,目前还不清楚控制 DNA 序列无关边界的组蛋白修饰是否以染色体特异性或非特异性的方式发挥作用。在这项研究中,我们评估了组蛋白乙酰转移酶(HAT)复合物 SAGA 复合物的作用及其与其他组蛋白修饰酶的关系,以阐明 VIII 号染色体右侧副染色体上 IMD2 基因的边界调控机制。我们发现,Spt8(SAGA 复合物的一个成分)对该区域的边界形成非常重要,而且 Spt8 加入 SAGA 复合物比其与 TATA 结合蛋白和 TFIIS 的相互作用更为重要。除 SAGA 外,NuA4 和 Rtt109 等多种 HAT 相关因子也在该区域发挥作用。特别是,SAGA 复合物诱导整个细胞中 IMD2 的弱表达,而 NuA4 则诱导强表达。这些结果表明,多种 HAT 参与了 VIII 号染色体右侧亚细胞组的边界形成和 IMD2 表达的调控,IMD2 的表达取决于这些因素之间的平衡。
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引用次数: 0
A novel tracking and analysis system for time-lapse cellular imaging of Schizosaccharomyces pombe. 用于对小鼠裂殖酵母进行延时细胞成像的新型跟踪和分析系统。
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-26 Epub Date: 2024-02-21 DOI: 10.1266/ggs.23-00239
Kei Taniguchi, Takuya Kajitani, Takahito Ayano, Toshiyuki Yoshida, Masaya Oki

The importance of the parent-progeny relationship tracking technique in single-cell analysis has grown with the passage of time. In this study, fundamental image-processing techniques were combined to develop software capable of inferring cell cycle alterations in fission yeast cells, which exhibit equipartition during division. These methods, exclusively relying on bright-field images as input, could track parent-progeny relationships after cell division by assessing the temporal morphological transformation of these cells. In the application of this technique, the software was employed for calculating intracellular fluorescent dots during every stage of the cell cycle, using a yeast strain expressing EGFP-fused Swi6, which binds to chromatin. The results obtained with this software were consistent with those of previous studies. This software facilitated single-cell-level tracking of parent-progeny relationships in cells exhibiting equipartition during division and enabled the monitoring of spatial fluctuations in a cell cycle-dependent protein. This method, expediting the analysis of extensive datasets, may also empower large-scale screening experiments that cannot be conducted manually.

随着时间的推移,在单细胞分析中采用亲子关系跟踪技术的意义越来越大。在这项研究中,我们综合运用了基本的图像处理技术,开发出了能够推断分裂酵母在分裂过程中细胞周期变化的软件。这些方法完全依赖于明视野图像作为输入,通过评估这些细胞的时间形态变化,可追踪细胞分裂后的亲子关系。在这项技术的应用中,利用酵母菌株 GFP 融合 Swi6(存在于细胞中并与染色质结合),使用软件计算细胞周期每个阶段的细胞内荧光点。该软件得出的结果与之前的研究结果一致。该软件有助于在单细胞水平上追踪分裂过程中表现出等分的细胞中的亲代-后代关系,并能监测依赖细胞周期的蛋白质的空间波动。这种方法加快了对大量数据集的分析,还可以进行人工无法进行的大规模筛选实验。
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引用次数: 0
Mouse retrotransposons: sequence structure, evolutionary age, genomic distribution and function. 小鼠反转录转座子:序列结构、进化年龄、基因组分布和功能。
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-10 Epub Date: 2023-11-22 DOI: 10.1266/ggs.23-00221
Masaki Kawase, Kenji Ichiyanagi

Retrotransposons are transposable elements that are transposed via transcription and reverse transcription. Their copies have accumulated in the genome of mammals, occupying approximately 40% of mammalian genomic mass. These copies are often involved in numerous phenomena, such as chromatin spatial organization, gene expression, development and disease, and have been recognized as a driving force in evolution. Different organisms have gained specific retrotransposon subfamilies and retrotransposed copies, such as hundreds of Mus-specific subfamilies with diverse sequences and genomic locations. Despite this complexity, basic information is still necessary for present-day genomic and epigenomic studies. Herein, we describe the characteristics of each subfamily of Mus-specific retrotransposons in terms of sequence structure, phylogenetic relationships, evolutionary age, and preference for A or B compartments of chromatin.

反转录转座子是通过转录和逆转录进行转座的转座元件。它们的拷贝在哺乳动物基因组中积累,占哺乳动物基因组质量的40%左右。这些拷贝经常参与许多现象,如染色质空间组织、基因表达、发育和疾病,并已被认为是进化的驱动力。不同的生物体已经获得了特定的反转录转座子亚家族和反转录转座子拷贝,例如数百个具有不同序列和基因组位置的小鼠特异性亚家族。尽管这种复杂性,基本信息仍然是必要的,今天的基因组和表观基因组的研究。在此,我们从序列结构、系统发育关系、进化年龄以及对染色质A或B区室的偏好等方面描述了每个鼠特异性反转录转座子亚家族的特征。
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引用次数: 0
Evolution of the nervous system by acquisition of retrovirus-derived genes in mammals. 哺乳动物通过获得逆转录病毒基因实现神经系统的进化。
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-10 Epub Date: 2024-01-13 DOI: 10.1266/ggs.23-00197
Moe Kitazawa

In the course of evolution, the most highly developed organ is likely the brain, which has become more complex over time and acquired diverse forms and functions in different species. In particular, mammals have developed complex and high-functioning brains, and it has been reported that several genes derived from retroviruses were involved in mammalian brain evolution, that is, generating the complexity of the nervous system. Especially, the sushi-ichi-related retrotransposon homolog (SIRH)/retrotransposon gag-like (RTL) genes have been suggested to play a role in the evolutionary processes shaping brain morphology and function in mammals. Genetic mutation and altered expression of genes are linked to neurological disorders, highlighting how the acquisition of virus-derived genes in mammals has both driven brain evolution and imposed a susceptibility to diseases. This review provides an overview of the functions, diversity, evolution and diseases associated with SIRH/RTL genes in the nervous system. The contribution of retroviruses to brain evolution is an important research topic in evolutionary biology and neuroscience, and further insights are expected to be gained through future studies.

在进化过程中,最发达的器官可能是大脑,随着时间的推移,大脑变得越来越复杂,在不同物种中获得了不同的形式和功能。尤其是哺乳动物,它们的大脑复杂而功能强大,有报道称,一些源自逆转录病毒的基因参与了哺乳动物大脑的进化,即产生了复杂的神经系统。特别是鮨一相关逆转录转座子同源物(SIRH)/类逆转录转座子gag(RTL)基因被认为在哺乳动物大脑形态和功能的进化过程中发挥了作用。基因突变和基因表达的改变与神经系统疾病有关,突显了哺乳动物体内病毒衍生基因的获得是如何推动大脑进化并导致易患疾病的。本综述概述了神经系统中 SIRH/RTL 基因的功能、多样性、进化和相关疾病。逆转录病毒对大脑进化的贡献是进化生物学和神经科学的一个重要研究课题,预计未来的研究将对此有更深入的了解。
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引用次数: 0
Complete mitochondrial and chloroplast DNA sequences of the freshwater green microalga Medakamo hakoo. 淡水绿色微藻 Medakamo hakoo 的完整线粒体和叶绿体 DNA 序列。
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-10 Epub Date: 2024-01-23 DOI: 10.1266/ggs.23-00275
Mari Takusagawa, Osami Misumi, Hisayoshi Nozaki, Shoichi Kato, Shinichiro Maruyama, Yayoi Tsujimoto-Inui, Fumi Yagisawa, Mio Ohnuma, Haruko Kuroiwa, Tsuneyoshi Kuroiwa, Sachihiro Matsunaga

We report the complete organellar genome sequences of an ultrasmall green alga, Medakamo hakoo strain M-hakoo 311, which has the smallest known nuclear genome in freshwater green algae. Medakamo hakoo has 90.8-kb chloroplast and 36.5-kb mitochondrial genomes containing 80 and 33 putative protein-coding genes, respectively. The mitochondrial genome is the smallest in the Trebouxiophyceae algae studied so far. The GC content of the nuclear genome is 73%, but those of chloroplast and mitochondrial genomes are 41% and 35%, respectively. Codon usages in the organellar genomes have a different tendency from that in the nuclear genome. The organellar genomes have unique characteristics, such as the biased encoding of mitochondrial genes on a single strand and the absence of operon structures in chloroplast ribosomal genes. Medakamo hakoo will be helpful for understanding the evolution of the organellar genome and the regulation of gene expression in chloroplasts and mitochondria.

我们报告了超小型绿藻 Medakamo hakoo 株 M-hakoo 311 的完整细胞器基因组序列,它是已知淡水绿藻中最小的核基因组。Medakamo hakoo 有 90.8 kb 的叶绿体基因组和 36.5 kb 的线粒体基因组,分别含有 80 和 33 个假定的蛋白质编码基因。线粒体基因组是迄今所研究的三叶藻类中最小的。核基因组的 GC 含量为 73%,而叶绿体和线粒体基因组的 GC 含量分别为 41% 和 35%。细胞器基因组中密码子的使用趋势与核基因组不同。细胞器基因组具有独特的特征,例如线粒体基因偏向于在单链上编码,叶绿体核糖体基因不存在操作子结构。Medakamo hakoo将有助于了解细胞器基因组的进化以及叶绿体和线粒体中基因表达的调控。
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引用次数: 0
Retrotransposon-derived transcripts and their functions in immunity and disease. 逆转录病毒载体产生的转录本及其在免疫和疾病中的功能
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-10 Epub Date: 2024-01-10 DOI: 10.1266/ggs.23-00187
Mahoko Takahashi Ueda

Retrotransposons, which account for approximately 42% of the human genome, have been increasingly recognized as "non-self" pathogen-associated molecular patterns (PAMPs) due to their virus-like sequences. In abnormal conditions such as cancer and viral infections, retrotransposons that are aberrantly expressed due to impaired epigenetic suppression display PAMPs, leading to their recognition by pattern recognition receptors (PRRs) of the innate immune system and triggering inflammation. This viral mimicry mechanism has been observed in various human diseases, including aging and autoimmune disorders. However, recent evidence suggests that retrotransposons possess highly regulated immune reactivity and play important roles in the development and function of the immune system. In this review, I discuss a wide range of retrotransposon-derived transcripts, their role as targets in immune recognition, and the diseases associated with retrotransposon activity. Furthermore, I explore the implications of chimeric transcripts formed between retrotransposons and known gene mRNAs, which have been previously underestimated, for the increase of immune-related gene isoforms and their influence on immune function. Retrotransposon-derived transcripts have profound and multifaceted effects on immune system function. The aim of this comprehensive review is to provide a better understanding of the complex relationship between retrotransposon transcripts and immune defense.

反转座子约占人类基因组的 42%,由于其类似病毒的序列,越来越多的人将其视为 "非自身 "病原体相关分子模式(PAMP)。在癌症和病毒感染等异常情况下,由于表观遗传抑制受损而异常表达的逆转录转座子会显示出 PAMPs,从而被先天性免疫系统的模式识别受体(PRRs)识别并引发炎症。在包括衰老和自身免疫性疾病在内的多种人类疾病中都观察到了这种病毒模仿机制。然而,最近的证据表明,逆转录病毒座子具有高度调节的免疫反应性,在免疫系统的发育和功能中发挥着重要作用。在这篇综述中,我将讨论一系列逆转录转座子衍生的转录本、它们在免疫识别中作为靶标的作用以及与逆转录转座子活性相关的疾病。此外,我还探讨了逆转录病毒载体与已知基因 mRNA 之间形成的嵌合转录本对免疫相关基因异构体的增加及其对免疫功能影响的影响,这些影响以前一直被低估。逆转录病毒座子衍生的转录本对免疫系统功能有深远和多方面的影响。本综述旨在让人们更好地了解逆转录病毒转录本与免疫防御之间的复杂关系。
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引用次数: 0
The role of transposable elements in human evolution and methods for their functional analysis: current status and future perspectives. 转座元件在人类进化中的作用及其功能分析方法:现状和未来展望。
IF 1.1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-10 Epub Date: 2023-10-20 DOI: 10.1266/ggs.23-00140
Kei Fukuda

Transposable elements (TEs) are mobile DNA sequences that can insert themselves into various locations within the genome, causing mutations that may provide advantages or disadvantages to individuals and species. The insertion of TEs can result in genetic variation that may affect a wide range of human traits including genetic disorders. Understanding the role of TEs in human biology is crucial for both evolutionary and medical research. This review discusses the involvement of TEs in human traits and disease susceptibility, as well as methods for functional analysis of TEs.

可转座元件(TE)是一种可移动的DNA序列,可以将其自身插入基因组内的不同位置,从而引起突变,从而为个体和物种提供优势或劣势。TE的插入可能导致遗传变异,这可能影响包括遗传疾病在内的广泛的人类特征。了解TE在人类生物学中的作用对于进化和医学研究都至关重要。这篇综述讨论了TEs与人类特征和疾病易感性的关系,以及TEs的功能分析方法。
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引用次数: 0
Author Index (vol. 99, 2024). 作者索引(第99卷,2024)。
IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-01-01 DOI: 10.1266/ggs.authorindex_99_2024
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引用次数: 0
Corrigendum: A new variant of the ectodysplasin A receptor death domain gene associated with anhidrotic ectodermal dysplasia in a Turkish family and its simple diagnosis by restriction fragment length polymorphism [Genes Genet. Syst. (2023) 98, p. 171-178]. 更正:一个土耳其家族中与湿疹性外胚层发育不良相关的外胚层发育蛋白 A 受体死亡域基因的新变异及其通过限制性片段长度多态性的简单诊断[基因遗传学系统(2023)98,第 171-178 页]。
IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-01-01 DOI: 10.1266/ggs.corrigendum_22-00138
Eyyup Rencuzogullari, Banu Guven Ezer

The ClinVar accession number on p. 176 (SCV002817173) should be replaced with the correct number, SCV002817373. The PDF file for DOI: https://doi.org/10.1266/ggs.22-00138 has been replaced with the corrected version as of November 6, 2024.

第 176 页的 ClinVar 编号(SCV002817173)应替换为正确的编号 SCV002817373。DOI: https://doi.org/10.1266/ggs.22-00138 的 PDF 文件已替换为 2024 年 11 月 6 日的更正版本。
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引用次数: 0
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Genes & genetic systems
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