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Identification of heparin-binding proteins expressed on Trichosporon asahii cell surface. 鉴定旭三代孢子虫细胞表面表达的肝素结合蛋白。
IF 2.2 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-01 Epub Date: 2024-01-31 DOI: 10.1002/yea.3928
Tomoe Ichikawa, Yuka Ikeda, Jumpei Sadanaga, Ayano Kikuchi, Kohei Kawamura, Reiko Ikeda, Yoshio Ishibashi

Trichosporon asahii is a pathogenic yeast that cause trichosporonosis. T. asahii exhibits several colony morphologies, such as white (W)- or off-white (O)-type, which may affect virulence. In this study, we compared the expression pattern of heparin-binding proteins in various colony morphologies and identified heparin-binding protein in T. asahii. Surface plasmon resonance analysis revealed that cell surface molecules attached more strongly to heparin in W- than O-type cells. We purified and identified a heparin-binding protein strongly expressed in W-type cells using heparin-Sepharose beads, named it heparin-binding protein 1 (HepBP1), and expressed Flag-tagged HepBP1 in mammalian cells. The heparin-binding ability of Flag-tagged HepBP1 was confirmed by pulldown assay using heparin-Sepharose beads. Thus, HepBP1 is a heparin-binding protein on T. asahii cell surface. These results suggest that several T. asahii cell surface proteins interact with glycosaminoglycans; therefore, they could contribute to infection.

Asahii 三孢酵母菌是一种致病酵母菌,可引起三孢酵母菌病。T. asahii表现出多种菌落形态,如白色(W)型或米白色(O)型,这可能会影响其毒力。在这项研究中,我们比较了肝素结合蛋白在不同菌落形态中的表达模式,并鉴定了 T. asahii 中的肝素结合蛋白。表面等离子共振分析表明,W 型细胞比 O 型细胞的细胞表面分子与肝素的附着力更强。我们利用肝素-Sepharose珠纯化并鉴定了一种在W型细胞中强表达的肝素结合蛋白,将其命名为肝素结合蛋白1(HepBP1),并在哺乳动物细胞中表达了Flag标记的HepBP1。利用肝素-Sepharose珠进行的pulldown试验证实了Flag标记的HepBP1的肝素结合能力。因此,HepBP1 是旭蚊细胞表面的肝素结合蛋白。这些结果表明,有几种麻风杆菌细胞表面蛋白与糖胺聚糖相互作用;因此,它们可能有助于感染。
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引用次数: 0
Yeast-insect interactions in southern Africa: Tapping the diversity of yeasts for modern bioprocessing. 南部非洲酵母与昆虫的相互作用:利用酵母的多样性进行现代生物加工。
IF 2.2 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-01 Epub Date: 2024-03-07 DOI: 10.1002/yea.3935
Tawanda P Makopa, Thembekile Ncube, Saleh Alwasel, Teun Boekhout, Nerve Zhou

Yeast-insect interactions are one of the most interesting long-standing relationships whose research has contributed to our understanding of yeast biodiversity and their industrial applications. Although insect-derived yeast strains are exploited for industrial fermentations, only a limited number of such applications has been documented. The search for novel yeasts from insects is attractive to augment the currently domesticated and commercialized production strains. More specifically, there is potential in tapping the insects native to southern Africa. Southern Africa is home to a disproportionately high fraction of global biodiversity with a cluster of biomes and a broad climate range. This review presents arguments on the roles of the mutualistic relationship between yeasts and insects, the presence of diverse pristine environments and a long history of spontaneous food and beverage fermentations as the potential source of novelty. The review further discusses the recent advances in novelty of industrial strains of insect origin, as well as various ancient and modern-day industries that could be improved by use yeasts from insect origin. The major focus of the review is on the relationship between insects and yeasts in southern African ecosystems as a potential source of novel industrial yeast strains for modern bioprocesses.

酵母与昆虫的相互作用是最有趣的长期关系之一,其研究有助于我们了解酵母的生物多样性及其工业应用。虽然昆虫衍生酵母菌株可用于工业发酵,但只有有限的此类应用记录在案。从昆虫中寻找新型酵母菌对增强目前的驯化和商业化生产菌株很有吸引力。更具体地说,挖掘南部非洲本地昆虫的潜力是巨大的。南部非洲的生物多样性在全球生物多样性中所占比例极高,拥有众多生物群落和广泛的气候范围。本综述论述了酵母菌与昆虫之间的互利关系、多样的原始环境和悠久的自发食品和饮料发酵历史作为潜在创新来源的作用。综述进一步讨论了昆虫源工业菌株新颖性的最新进展,以及可通过使用昆虫源酵母改善的各种古代和现代工业。综述的主要重点是南部非洲生态系统中昆虫与酵母菌之间的关系,它们是现代生物工艺中新型工业酵母菌株的潜在来源。
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引用次数: 0
Yeast Crf1p is an activator with different roles in regulation of target genes 酵母 Crf1p 是一种激活剂,在靶基因调控中发挥不同作用
IF 2.6 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-19 DOI: 10.1002/yea.3939
Sanjay Kumar, Muneera Mashkoor, Priya Balamurugan, Anne Grove
Under stress conditions, ribosome biogenesis is downregulated. This process requires that expression of ribosomal RNA, ribosomal protein, and ribosome biogenesis genes be controlled in a coordinated fashion. The mechanistic Target of Rapamycin Complex 1 (mTORC1) participates in sensing unfavorable conditions to effect the requisite change in gene expression. In Saccharomyces cerevisiae, downregulation of ribosomal protein genes involves dissociation of the activator Ifh1p in a process that depends on Utp22p, a protein that also functions in pre‐rRNA processing. Ifh1p has a paralog, Crf1p, which was implicated in communicating mTORC1 inhibition and hence was perceived as a repressor. We focus here on two ribosomal biogenesis genes, encoding Utp22p and the high mobility group protein Hmo1p, both of which are required for communication of mTORC1 inhibition to target genes. Crf1p functions as an activator on these genes as evidenced by reduced mRNA abundance and RNA polymerase II occupancy in a crf1Δ strain. Inhibition of mTORC1 has distinct effects on expression of HMO1 and UTP22; for example, on UTP22, but not on HMO1, the presence of Crf1p promotes the stable depletion of Ifh1p. Our data suggest that Crf1p functions as a weak activator, and that it may be required to prevent re‐binding of Ifh1p to some gene promoters after mTORC1 inhibition in situations when Ifh1p is available. We propose that the inclusion of genes encoding proteins required for mTORC1‐mediated downregulation of ribosomal protein genes in the same regulatory circuit as the ribosomal protein genes serves to optimize transcriptional responses during mTORC1 inhibition.
在应激条件下,核糖体生物发生受到下调。这一过程要求以协调的方式控制核糖体 RNA、核糖体蛋白和核糖体生物发生基因的表达。雷帕霉素复合体 1 的机制靶标(mTORC1)参与感知不利条件,以实现基因表达的必要变化。在酿酒酵母(Saccharomyces cerevisiae)中,核糖体蛋白基因的下调涉及激活剂 Ifh1p 的解离过程,这一过程依赖于 Utp22p,后者也是一种在前 RNA 处理中发挥作用的蛋白质。Ifh1p 有一个旁系亲属 Crf1p,它与 mTORC1 的抑制作用有关,因此被认为是一种抑制因子。我们在此重点研究两个核糖体生物发生基因,它们分别编码 Utp22p 和高迁移率基团蛋白 Hmo1p,这两个基因都是将 mTORC1 抑制作用传递给靶基因所必需的。Crf1p 可作为这些基因的激活剂,这一点可从 crf1Δ 菌株中减少的 mRNA 丰度和 RNA 聚合酶 II 占有率得到证明。抑制 mTORC1 会对 HMO1 和 UTP22 的表达产生不同的影响;例如,Crf1p 的存在会促进 Ifh1p 的稳定消耗,但对 UTP22 而言却不会对 HMO1 产生影响。我们的数据表明,Crf1p 起着弱激活剂的作用,在 Ifh1p 可用的情况下,可能需要它来防止 mTORC1 抑制后 Ifh1p 与某些基因启动子重新结合。我们提出,将编码 mTORC1 介导的核糖体蛋白基因下调所需蛋白的基因与核糖体蛋白基因纳入同一调控回路,可优化 mTORC1 抑制期间的转录反应。
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引用次数: 0
Comparative transcriptome analysis reveals the redirection of metabolic flux from cell growth to astaxanthin biosynthesis in Yarrowia lipolytica 比较转录组分析揭示了脂肪溶解亚罗维氏菌中从细胞生长到虾青素生物合成的代谢通量的重新定向
IF 2.6 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-13 DOI: 10.1002/yea.3938
Dan‐Ni Wang, Chen‐Xi Yu, Jie Feng, Liu‐Jing Wei, Jun Chen, Zhijie Liu, Liming Ouyang, Lixin Zhang, Feng Liu, Qiang Hua
Engineering Yarrowia lipolytica to produce astaxanthin provides a promising route. Here, Y. lipolytica M2 producing a titer of 181 mg/L astaxanthin was isolated by iterative atmospheric and room‐temperature plasma mutagenesis and diphenylamine‐mediated screening. Interestingly, a negative correlation was observed between cell biomass and astaxanthin production. To reveal the underlying mechanism, RNA‐seq analysis of transcriptional changes was performed in high producer M2 and reference strain M1, and a total of 1379 differentially expressed genes were obtained. Data analysis revealed that carbon flux was elevated through lipid metabolism, acetyl‐CoA and mevalonate supply, but restrained through central carbon metabolism in strain M2. Moreover, upregulation of other pathways such as ATP‐binding cassette transporter and thiamine pyrophosphate possibly provided more cofactors for carotenoid hydroxylase and relieved cell membrane stress caused by astaxanthin insertion. These results suggest that balancing cell growth and astaxanthin production may be important to promote efficient biosynthesis of astaxanthin in Y. lipolytica.
对脂肪溶解蓍草菌进行工程改造以生产虾青素是一条很有前景的途径。在这里,通过大气和室温等离子体诱变迭代以及二苯胺介导的筛选,分离出了可产生 181 毫克/升虾青素滴度的脂溶性酵母菌 M2。有趣的是,细胞生物量与虾青素产量之间呈负相关。为揭示其潜在机制,对高产菌株 M2 和参考菌株 M1 的转录变化进行了 RNA-seq 分析,共获得 1379 个差异表达基因。数据分析显示,在菌株 M2 中,碳通量通过脂质代谢、乙酰-CoA 和甲羟戊酸供应得到提高,但通过中心碳代谢受到抑制。此外,ATP 结合盒转运体和焦磷酸硫胺素等其他途径的上调可能为类胡萝卜素羟化酶提供了更多辅助因子,并缓解了虾青素插入造成的细胞膜压力。这些结果表明,平衡细胞生长和虾青素生产可能对促进溶脂酵母中虾青素的高效生物合成非常重要。
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引用次数: 0
Gene transcription in yeasts: From molecules to integrated processes 酵母中的基因转录:从分子到综合过程
IF 2.6 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-08 DOI: 10.1002/yea.3936
Domenico Libri, Jane Mellor, Françoise Stutz, Benoit Palancade
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引用次数: 0
Live‐cell fluorescence imaging and optogenetic control of PKA kinase activity in fission yeast Schizosaccharomyces pombe 裂殖酵母中 PKA 激酶活性的活细胞荧光成像和光遗传控制
IF 2.6 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-07 DOI: 10.1002/yea.3937
Keiichiro Sakai, Kazuhiro Aoki, Yuhei Goto
The cAMP‐PKA signaling pathway plays a crucial role in sensing and responding to nutrient availability in the fission yeast Schizosaccharomyces pombe. This pathway monitors external glucose levels to control cell growth and sexual differentiation. However, the temporal dynamics of the cAMP‐PKA pathway in response to external stimuli remains unclear mainly due to the lack of tools to quantitatively visualize the activity of the pathway. Here, we report the development of the kinase translocation reporter (KTR)‐based biosensor spPKA‐KTR1.0, which allows us to measure the dynamics of PKA activity in fission yeast cells. The spPKA‐KTR1.0 is derived from the transcription factor Rst2, which translocates from the nucleus to the cytoplasm upon PKA activation. We found that spPKA‐KTR1.0 translocates between the nucleus and cytoplasm in a cAMP‐PKA pathway‐dependent manner, indicating that the spPKA‐KTR1.0 is a reliable indicator of the PKA activity in fission yeast cells. In addition, we implemented a system that simultaneously visualizes and manipulates the cAMP‐PKA signaling dynamics by introducing bPAC, a photoactivatable adenylate cyclase, in combination with spPKA‐KTR1.0. This system offers an opportunity for investigating the role of the signaling dynamics of the cAMP‐PKA pathway in fission yeast cells with higher temporal resolution.
cAMP-PKA 信号通路在裂殖酵母(Schizosaccharomyces pombe)中感知和响应营养供应方面起着至关重要的作用。该通路监测外部葡萄糖水平,以控制细胞生长和性分化。然而,cAMP-PKA 通路响应外部刺激的时间动态仍不清楚,这主要是因为缺乏定量可视化该通路活性的工具。在这里,我们报告了基于激酶转位报告(KTR)的生物传感器 spPKA-KTR1.0 的开发情况,它使我们能够测量裂殖酵母细胞中 PKA 活性的动态。spPKA-KTR1.0 源自转录因子 Rst2,它在 PKA 激活时会从细胞核转位到细胞质。我们发现,spPKA-KTR1.0在细胞核和细胞质之间的转位是依赖于cAMP-PKA途径的,这表明spPKA-KTR1.0是裂殖酵母细胞中PKA活性的可靠指标。此外,我们还通过引入可光激活的腺苷酸环化酶 bPAC 和 spPKA-KTR1.0,建立了一个可同时观察和操纵 cAMP-PKA 信号动态的系统。该系统为以更高的时间分辨率研究裂殖酵母细胞中 cAMP-PKA 通路信号动态的作用提供了机会。
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引用次数: 0
Transcription as source of genetic heterogeneity in budding yeast. 转录是芽殖酵母遗传异质性的来源。
IF 2.2 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-01 Epub Date: 2024-01-09 DOI: 10.1002/yea.3926
Baptiste Piguet, Jonathan Houseley

Transcription presents challenges to genome stability both directly, by altering genome topology and exposing single-stranded DNA to chemical insults and nucleases, and indirectly by introducing obstacles to the DNA replication machinery. Such obstacles include the RNA polymerase holoenzyme itself, DNA-bound regulatory factors, G-quadruplexes and RNA-DNA hybrid structures known as R-loops. Here, we review the detrimental impacts of transcription on genome stability in budding yeast, as well as the mitigating effects of transcription-coupled nucleotide excision repair and of systems that maintain DNA replication fork processivity and integrity. Interactions between DNA replication and transcription have particular potential to induce mutation and structural variation, but we conclude that such interactions must have only minor effects on DNA replication by the replisome with little if any direct mutagenic outcome. However, transcription can significantly impair the fidelity of replication fork rescue mechanisms, particularly Break Induced Replication, which is used to restart collapsed replication forks when other means fail. This leads to de novo mutations, structural variation and extrachromosomal circular DNA formation that contribute to genetic heterogeneity, but only under particular conditions and in particular genetic contexts, ensuring that the bulk of the genome remains extremely stable despite the seemingly frequent interactions between transcription and DNA replication.

转录会直接改变基因组拓扑结构,并使单链 DNA 遭受化学物质和核酸酶的侵蚀,从而对基因组的稳定性构成挑战;转录也会间接给 DNA 复制机制带来障碍。这些障碍包括 RNA 聚合酶全酶本身、与 DNA 结合的调控因子、G-四链体和称为 R 环的 RNA-DNA 杂交结构。在此,我们回顾了转录对芽殖酵母基因组稳定性的不利影响,以及转录耦合核苷酸切除修复和维持 DNA 复制叉过程性和完整性的系统的缓解作用。DNA 复制与转录之间的相互作用特别有可能诱发突变和结构变异,但我们的结论是,这种相互作用肯定只会对复制体的 DNA 复制产生轻微影响,几乎不会产生直接的诱变结果。然而,转录会极大地损害复制叉挽救机制的保真度,尤其是断裂诱导复制(Break Induced Replication)。这会导致新的突变、结构变异和染色体外环状 DNA 的形成,从而造成遗传异质性,但只有在特定的条件和特定的遗传背景下才会出现这种情况,从而确保尽管转录和 DNA 复制之间似乎经常发生相互作用,但基因组的大部分仍然非常稳定。
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引用次数: 0
Budding yeast as an ideal model for elucidating the role of N6-methyladenosine in regulating gene expression. 芽殖酵母是阐明 N6 -甲基腺苷调控基因表达作用的理想模型。
IF 2.2 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-01 Epub Date: 2024-01-18 DOI: 10.1002/yea.3925
Waleed S Albihlal, Wei Yee Chan, Folkert J van Werven

N6-methyladenosine (m6A) is a highly abundant and evolutionarily conserved messenger RNA (mRNA) modification. This modification is installed on RRACH motifs on mRNAs by a hetero-multimeric holoenzyme known as m6A methyltransferase complex (MTC). The m6A mark is then recognised by a group of conserved proteins known as the YTH domain family proteins which guide the mRNA for subsequent downstream processes that determine its fate. In yeast, m6A is installed on thousands of mRNAs during early meiosis by a conserved MTC and the m6A-modified mRNAs are read by the YTH domain-containing protein Mrb1/Pho92. In this review, we aim to delve into the recent advances in our understanding of the regulation and roles of m6A in yeast meiosis. We will discuss the potential functions of m6A in mRNA translation and decay, unravelling their significance in regulating gene expression. We propose that yeast serves as an exceptional model organism for the study of fundamental molecular mechanisms related to the function and regulation of m6A-modified mRNAs. The insights gained from yeast research not only expand our knowledge of mRNA modifications and their molecular roles but also offer valuable insights into the broader landscape of eukaryotic posttranscriptional regulation of gene expression.

N6 -甲基腺苷(m6A)是一种高度丰富和进化保守的信使 RNA(mRNA)修饰。这种修饰由一种称为 m6A 甲基转移酶复合物(MTC)的异源多聚体全酶安装在 mRNA 上的 RRACH 基序上。m6A 标记随后会被一组称为 YTH 结构域家族蛋白的保守蛋白识别,这些蛋白会引导 mRNA 进入决定其命运的后续下游过程。在酵母中,m6A 在减数分裂早期被一个保守的 MTC 安装到成千上万的 mRNA 上,m6A 修饰的 mRNA 被含 YTH 结构域的蛋白 Mrb1/Pho92 读取。在这篇综述中,我们将深入探讨我们对 m6A 在酵母减数分裂过程中的调控和作用的最新理解进展。我们将讨论 m6A 在 mRNA 翻译和衰变中的潜在功能,揭示它们在调控基因表达中的意义。我们认为,酵母是研究与 m6A 修饰的 mRNA 的功能和调控有关的基本分子机制的特殊模式生物。从酵母研究中获得的启示不仅拓展了我们对 mRNA 修饰及其分子作用的认识,而且为我们了解真核生物转录后基因表达调控的更广阔领域提供了宝贵的见解。
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引用次数: 0
How is polyadenylation restricted to 3'-untranslated regions? 聚腺苷化如何局限于3'-非翻译区?
IF 2.2 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-01 Epub Date: 2023-12-02 DOI: 10.1002/yea.3915
Kevin Struhl

Polyadenylation occurs at numerous sites within 3'-untranslated regions (3'-UTRs) but rarely within coding regions. How does Pol II travel through long coding regions without generating poly(A) sites, yet then permits promiscuous polyadenylation once it reaches the 3'-UTR? The cleavage/polyadenylation (CpA) machinery preferentially associates with 3'-UTRs, but it is unknown how its recruitment is restricted to 3'-UTRs during Pol II elongation. Unlike coding regions, 3'-UTRs have long AT-rich stretches of DNA that may be important for restricting polyadenylation to 3'-UTRs. Recognition of the 3'-UTR could occur at the DNA (AT-rich), RNA (AU-rich), or RNA:DNA hybrid (rU:dA- and/or rA:dT-rich) level. Based on the nucleic acid critical for 3'-UTR recognition, there are three classes of models, not mutually exclusive, for how the CpA machinery is selectively recruited to 3'-UTRs, thereby restricting where polyadenylation occurs: (1) RNA-based models suggest that the CpA complex directly (or indirectly through one or more intermediary proteins) binds long AU-rich stretches that are exposed after Pol II passes through these regions. (2) DNA-based models suggest that the AT-rich sequence affects nucleosome depletion or the elongating Pol II machinery, resulting in dissociation of some elongation factors and subsequent recruitment of the CpA machinery. (3) RNA:DNA hybrid models suggest that preferential destabilization of the Pol II elongation complex at rU:dA- and/or rA:dT-rich duplexes bridging the nucleotide addition and RNA exit sites permits preferential association of the CpA machinery with 3'-UTRs. Experiments to provide evidence for one or more of these models are suggested.

聚腺苷酸化发生在3'-非翻译区(3'- utr)的许多位点,但很少发生在编码区。Pol II如何穿过长编码区而不产生聚(A)位点,然而一旦到达3'-UTR就允许混杂聚腺苷化?切割/聚腺苷化(CpA)机制优先与3'- utr相关,但在Pol II延伸期间,其招募如何仅限于3'- utr尚不清楚。与编码区不同,3'- utr具有很长的富含at的DNA延伸,这对于限制3'- utr的聚腺苷化可能很重要。3'- utr的识别可能发生在DNA(富含at), RNA(富含au)或RNA:DNA杂交(rU:dA-和/或rA: dt -丰富)水平。基于对3’-UTR识别至关重要的核酸,对于CpA机制如何选择性地招募到3’-UTR,从而限制聚腺苷化发生的位置,有三种并非相互排斥的模型:(1)基于rna的模型表明,CpA复合体直接(或间接通过一个或多个中间蛋白)结合富含au的长片段,这些片段在Pol II通过这些区域后暴露。(2)基于dna的模型表明,富含at的序列影响核小体耗竭或延长Pol II机制,导致一些延长因子的解离和随后的CpA机制的招募。(3) RNA:DNA杂交模型表明,连接核苷酸加成位点和RNA退出位点的rU:dA-和/或rA: dt -富双链上Pol II延伸复合物的优先失稳,允许CpA机制与3'- utr优先结合。建议进行实验,为其中一个或多个模型提供证据。
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引用次数: 0
Size fractionated NET-Seq reveals a conserved architecture of transcription units around yeast genes. 大小分馏NET-Seq揭示了酵母基因周围转录单元的保守结构。
IF 2.2 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-04-01 Epub Date: 2024-03-03 DOI: 10.1002/yea.3931
Shidong Xi, Tania Nguyen, Struan Murray, Phil Lorenz, Jane Mellor

Genomes from yeast to humans are subject to pervasive transcription. A single round of pervasive transcription is sufficient to alter local chromatin conformation, nucleosome dynamics and gene expression, but is hard to distinguish from background signals. Size fractionated native elongating transcript sequencing (sfNET-Seq) was developed to precisely map nascent transcripts independent of expression levels. RNAPII-associated nascent transcripts are fractionation into different size ranges before library construction. When anchored to the transcription start sites (TSS) of annotated genes, the combined pattern of the output metagenes gives the expected reference pattern. Bioinformatic pattern matching to the reference pattern identified 9542 transcription units in Saccharomyces cerevisiae, of which 47% are coding and 53% are noncoding. In total, 3113 (33%) are unannotated noncoding transcription units. Anchoring all transcription units to the TSS or polyadenylation site (PAS) of annotated genes reveals distinctive architectures of linked pairs of divergent transcripts approximately 200nt apart. The Reb1 transcription factor is enriched 30nt downstream of the PAS only when an upstream (TSS -60nt with respect to PAS) noncoding transcription unit co-occurs with a downstream (TSS +150nt) coding transcription unit and acts to limit levels of upstream antisense transcripts. The potential for extensive transcriptional interference is evident from low abundance unannotated transcription units with variable TSS (median -240nt) initiating within a 500nt window upstream of, and transcribing over, the promoters of protein-coding genes. This study confirms a highly interleaved yeast genome with different types of transcription units altering the chromatin landscape in distinctive ways, with the potential to exert extensive regulatory control.

从酵母到人类的基因组都受到普遍转录的影响。一轮普遍转录足以改变局部染色质构象、核小体动力学和基因表达,但很难从背景信号中区分出来。我们开发了尺寸分馏原生延伸转录本测序(sfNET-Seq),以精确绘制新生转录本的图谱,而不受表达水平的影响。在构建文库之前,先将 RNAPII 相关的新生转录本分成不同的大小范围。当锚定到注释基因的转录起始位点(TSS)时,输出元基因的组合模式会给出预期的参考模式。与参考模式匹配的生物信息学模式在酿酒酵母中发现了 9542 个转录单元,其中 47% 是编码单元,53% 是非编码单元。总共有 3113 个(33%)是未注释的非编码转录单位。将所有转录单元锚定到已注释基因的 TSS 或多聚腺苷酸化位点(PAS),可发现相距约 200nt 的不同转录本成对连接的独特结构。只有当上游(TSS -60nt,相对于 PAS)非编码转录单位与下游(TSS +150nt )编码转录单位共存时,Reb1 转录因子才会在 PAS 下游 30nt 处富集,并限制上游反义转录本的水平。在蛋白编码基因启动子上游 500nt 窗口内启动并转录超过蛋白编码基因启动子的低丰度未注释转录单元(TSS 中位数为 -240nt)明显具有广泛转录干扰的潜力。这项研究证实,酵母基因组高度交错,不同类型的转录单位以独特的方式改变染色质景观,并有可能发挥广泛的调控作用。
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引用次数: 0
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