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Interrogation of the cell wall integrity pathway in Aspergillus niger identifies a putative negative regulator of transcription involved in chitin deposition. 对黑曲霉细胞壁完整性途径的研究发现了一种参与几丁质沉积的转录负调控因子。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2020-01-28 eCollection Date: 2020-12-01 DOI: 10.1016/j.gene.2020.100028
Tim M van Leeuwe, Mark Arentshorst, Peter J Punt, Arthur F J Ram

Post-fermentation fungal biomass waste provides a viable source for chitin. Cell wall chitin of filamentous fungi, and in particular its de-N-acetylated derivative chitosan, has a wide range of commercial applications. Although the cell wall of filamentous fungi comprises 10-30% chitin, these yields are too low for cost-effective production. Therefore, we aimed to identify the genes involved in increased chitin deposition by screening a collection of UV-derived cell wall mutants in Aspergillus niger. This screen revealed a mutant strain (RD15.4#55) that showed a 30-40% increase in cell wall chitin compared to the wild type. In addition to the cell wall chitin phenotype, this strain also exhibited sensitivity to SDS and produces an unknown yellow pigment. Genome sequencing combined with classical genetic linkage analysis identified two mutated genes on chromosome VII that were linked with the mutant phenotype. Single gene knockouts and subsequent complementation analysis revealed that an 8 bp deletion in NRRL3_09595 is solely responsible for the associated phenotypes of RD15.4#55. The mutated gene, which was named cwcA (cell wall chitin A), encodes an orthologue of Saccharomyces cerevisiae Bypass of ESS1 (BYE1), a negative regulator of transcription elongation. We propose that this conserved fungal protein is involved in preventing cell wall integrity signaling under non-inducing conditions, where loss of function results in constitutive activation of the cell wall stress response pathway, and consequently leads to increased chitin content in the mutant cell wall.

发酵后的真菌生物质废物为甲壳素提供了可行的来源。丝状真菌细胞壁甲壳素,特别是其去 N-乙酰化衍生物壳聚糖,具有广泛的商业用途。虽然丝状真菌的细胞壁含有 10-30% 的几丁质,但这些产量太低,无法进行具有成本效益的生产。因此,我们筛选了一系列黑曲霉的紫外线衍生细胞壁突变体,旨在确定参与增加几丁质沉积的基因。筛选结果显示,与野生型相比,突变株(RD15.4#55)的细胞壁几丁质增加了 30-40%。除了细胞壁几丁质表型外,该菌株还表现出对 SDS 的敏感性,并产生一种未知的黄色色素。基因组测序与经典遗传连锁分析相结合,在 VII 号染色体上发现了两个与突变表型相关的突变基因。单基因敲除和随后的互补分析表明,NRRL3_09595 中一个 8 bp 的缺失是造成 RD15.4#55 相关表型的唯一原因。突变基因被命名为 cwcA(细胞壁甲壳素 A),它编码酿酒酵母ESS1 的旁路(BYE1)的同源物,后者是转录伸长的负调控因子。我们认为这种保守的真菌蛋白参与了非诱导条件下细胞壁完整性信号的阻止,功能缺失会导致细胞壁应激反应途径的组成性激活,从而导致突变体细胞壁中几丁质含量的增加。
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引用次数: 0
Putative circumsporozoite protein (CSP) of Plasmodium vivax is considerably distinct from the well-known CSP and plays a role in the protein ubiquitination pathway. 间日疟原虫的假定环孢子虫蛋白(CSP)与众所周知的 CSP 有很大不同,它在蛋白质泛素化途径中发挥作用。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-11-12 eCollection Date: 2019-12-01 DOI: 10.1016/j.gene.2019.100024
Manoswini Dash, Veena Pande, Abhinav Sinha

Amidst technical challenges which limit successful culture and genetic manipulation of P. vivax parasites, we used a computational approach to identify a critical target with evolutionary significance. The putative circumsporozoite protein on chromosome 13 of P. vivax (PvpuCSP)is distinct from the well-known vaccine candidate PfCSP. The aim of this study was to understand the role of PvpuCSP and its relatedness to the well-known CSP. The study revealed PvpuCSP as a membrane bound E3 ubiquitin ligase involved in ubiquitination. It has a species-specific tetra-peptide unit which is differentially repeated in various P. vivax strains. The PvpuCSP is different from CSP in terms of stage-specific expression and function. Since E3 ubiquitin ligases are known antimalarial drug targets targeting the proteasome pathway, PvpuCSP, with evolutionary connotation and a key role in orchestrating protein degradation in P. vivax, can be explored as a potential drug target.

技术上的挑战限制了对间鞭毛虫寄生虫的成功培养和基因操作,在这种情况下,我们利用计算方法确定了一个具有进化意义的关键靶标。间日疟原虫第13号染色体上的假定环孢子虫蛋白(PvpuCSP)与众所周知的候选疫苗PfCSP不同。本研究的目的是了解 PvpuCSP 的作用及其与众所周知的 CSP 的关系。研究发现,PvpuCSP 是一种膜结合 E3 泛素连接酶,参与泛素化。它有一个物种特异性的四肽单元,在不同的间日疟原虫菌株中有不同的重复。PvpuCSP 在特异性表达和功能方面与 CSP 不同。由于 E3 泛素连接酶是针对蛋白酶体途径的已知抗疟疾药物靶标,PvpuCSP 具有进化内涵,在协调间日疟原虫蛋白质降解方面发挥着关键作用,因此可作为潜在的药物靶标进行研究。
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引用次数: 0
Upregulation of the long non-coding RNA CBR3-AS1 predicts tumor prognosis and contributes to breast cancer progression. 长非编码 RNA CBR3-AS1 的上调可预测肿瘤的预后,并有助于乳腺癌的进展。
Q3 Biochemistry, Genetics and Molecular Biology Pub Date : 2019-03-25 eCollection Date: 2019-06-01 DOI: 10.1016/j.gene.2019.100014
Lingyun Xu, Hong Zhu, Fei Gao, Yinghua Tang, Yajun Zhu, Zhiqiang Sun, Jian Wang

Breast cancer is the most common female malignancy and the major cause of cancer-related death in women. Long non-coding RNAs (lncRNAs), as oncogenic or tumor suppressor factor, involved in the development and progression of various cancers. In this study, we sought to investigate the function of lncRNA CBR3-AS1 in breast cancer. We evaluated the expression pattern of CBR3-AS1 in breast cancer tissues and cell lines, explored the correlation between CBR3-AS1 expression and the survival time of breast cancer patients, and probed the effect of CBR3-AS1 on tumor progression of breast cancer through loss-of-function and gain-of-function strategies. Our results showed that CBR3-AS1 was overexpressed in breast cancer tissues and cell lines and predicted the prognosis of breast cancer patients. And CBR3-AS1 exerted biological function as an oncogenic lncRNA, involved in the regulation of cell proliferation, colony formation, apoptosis and tumor growth in breast cancer. Taken together, CBR3-AS1 was up-regulated in breast cancer and promoted the risk of breast cancer. It may be a novel therapeutic target and potential prognostic marker for breast cancer.

乳腺癌是最常见的女性恶性肿瘤,也是女性因癌症死亡的主要原因。长非编码 RNA(lncRNA)作为致癌或抑癌因子,参与了多种癌症的发生和发展。在这项研究中,我们试图研究lncRNA CBR3-AS1在乳腺癌中的功能。我们评估了CBR3-AS1在乳腺癌组织和细胞系中的表达模式,探讨了CBR3-AS1表达与乳腺癌患者生存时间的相关性,并通过功能缺失和功能获得策略探讨了CBR3-AS1对乳腺癌肿瘤进展的影响。结果表明,CBR3-AS1在乳腺癌组织和细胞系中过表达,可预测乳腺癌患者的预后。CBR3-AS1作为一种致癌lncRNA,参与调控乳腺癌的细胞增殖、集落形成、细胞凋亡和肿瘤生长。综上所述,CBR3-AS1 在乳腺癌中上调,并增加了乳腺癌的风险。它可能是乳腺癌的一个新的治疗靶点和潜在的预后标志物。
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引用次数: 0
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Gene: X
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