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Apoptotic endonuclease EndoG plays a key role in the alternative splicing of mRNA of human TERT telomerase catalytic subunit. The aim of this work was to test the ability of EndoG to induce alternative splicing of mRNA of other genes and in other organisms. To determine new mRNA splice-variants, EndoG overexpression was induced in human, mouse and rat CD4+-T-lymphocytes followed by sequencing of total RNA of these cells. Sequencing results showed that besides TERT, EndoG induced alternative splicing of deoxyribonuclease I (DNase I), caspase-2 (Casp-2) and BCL-x. The expression level of EndoG strongly correlated with mRNA splicing-variants of TERT, DNase I, Casp-2, and BCL-x in intact CD4+-T cells of healthy donors as well as different lines of mice and rats. EndoG overexpression induced down-regulation of fulllength mRNAs of TERT, DNase I, Casp-2, and BCL-x and up-regulation of their short-length mRNAs. Alternative splicing of studied mRNAs resulted in down-regulation of enzymatic activity of proteins in vitro and in vivo. The results of this work confirm the ability of endonuclease EndoG to induce alternative splicing of several mRNAs in human, mice and rats. 相似文献
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用 RT-PCR 引物分别扩增成年昆明 (KM) 小鼠睾丸、脾脏、肾脏、肝脏和胸腺组织的总 RNA 发现,端粒酶催化亚基基因 tert 在这些组织中都有转录,目标产物正确组装到 PMD 18-T 载体后测序,结果与已知 cDNA 序列一致 . PMSG/hCG 超数排卵方法获得 KM 小鼠成熟卵母细胞和 CZB 溶液体外培养的胚胎 (KM ♀× KM ♂ ) ,用酸性 Tyrode's 溶液消化透明带后,采用巢式 RT-PCR ,同时分析 tert 基因和持家基因 hprt 的转录发现,对于单个样品来说 , 全部卵母细胞 (15 h-post hCG , 10/10) 都存在 hprt 转录本,其中,只有 40% (4/10) 还同时存在 tert 转录本 . 原核形成初期 (20 h-post hCG , 6/6) 和原核晚期 (30 h post-hCG , 8/8) 的受精卵,以及发育至 2-C 早期的胚胎 (35 h-post hCG , 7/7) 都不转录 tert 基因,只有 hprt mRNA 存在; 2-C 晚期 (50 h-post hCG) 时,两个基因同时转录 (4/8) 和一个基因单独转录 (4/8) 的胚胎各占 50% ;从 4-C 阶段 (65 h-post hCG , 4/4) 开始,包括 8-C 阶段 (75 h-post hCG , 4/4) ,桑椹胚阶段 (93 h-post hCG , 4/4) ,直至囊胚阶段 (118 h-post hCG , 4/4) ,所有的胚胎都同时转录 tert 和 hprt 基因,而且转录水平明显升高 . 以 20 枚胚胎量为模板进行 RT-PCR 发现,原核早期,原核晚期的胚胎中仍然没有 tert 基因转录,只有 hprt mRNA ,但是,在 2-C 早期胚胎中同时检测到了 hprt 和 tert 两种 mRNA. 结果表明,持家基因 hprt 在成熟卵母细胞受精前后,以及胚胎早期发育过程中均存在转录本 . 40% 卵母细胞中存在的 tert mRNA 在受精后很快降解,检测不到;胚胎基因组在 2-C 早期开始转录 tert mRNA ,转录水平逐渐上升 . 结果暗示,小鼠胚胎的基因组 DNA 在 2-C 早期开始启动,功能基因 tert 也在此时开始转录,可能与胚胎发育初期的染色体保护有关 . 相似文献
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《Cell cycle (Georgetown, Tex.)》2013,12(12):1286-1288
It has long been suggested that alternative splicing is involved in regulation of apoptosis by producing mRNA isoforms that encode proteins with distinct and even opposite functions in apoptotic pathways. However, the physiological functions and regulatory mechanisms of such alternative splicing events have been unclear. Recently, it was demonstrated that inactivation of a single SR protein, ASF/SF2, can modulate a specific step in the apoptotic pathway, internucleosomal DNA fragmentation, by regulating ICAD pre-mRNA alternative splicing. These studies have provided new evidence supporting the important role of regulated splicing and SR proteins in the process of apoptosis. 相似文献
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Mandy S. Wong Ling Chen Christopher Foster Radhika Kainthla Jerry W. Shay Woodring E. Wright 《Cell reports》2013,3(4):1028-1035
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Campusano JM Andrés ME Magendzo K Abarca J Tapia-Arancibia L Bustos G 《Neurochemical research》2005,30(4):567-576
The expression of mesencephalic brain derived neurotrophic factor (BDNF) has been shown to be regulated by dopaminergic neuronal functioning and glutamate receptors (GluRs). In turn, BDNF participates in the regulation of mesencephalic GluRs’ expression. In the present study we analyzed, using semi-quantitative RT-PCR, the effect of BDNF as well as of the GluRs agonists NMDA and trans-(±)-1-Amino-(1S,3R)-cyclopentane dicarboxylic acid (t-ACPD), on the expression levels of the NMDA GluR subunit 1 (NR1) mRNA, using rat cultured mesencephalic neurons. In the course of this study, a novel rat mRNA splice variant of NR1 was identified. This new NR1 mRNA isoform is characterized by the insertion of an 82 base pair intron containing an inframe stop codon, thus predicting the expression of a putative truncated protein of 465 amino acids. The RT-PCR and in situ hybridization reveals that the novel NR1 mRNA is expressed in various brain regions of the rat embryo, whereas no expression was detected in the adult rat brain. The modulation of the novel NR1 mRNA isoform by both BDNF and the metabotropic GluR agonist t-ACPD, suggests that the resulting putative NR1 truncated protein may be relevant in the regulatory network of glutamatergic neurotransmission in the developing central nervous system. 相似文献
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端粒酶是一种核糖核蛋白复合物 ,能引起染色体的末端结构端粒的完全复制。端粒作为一种保护性结构 ,是由短的重复DNA序列组成。在人体中这种序列为TTAGGG ,其平均长度为 5~ 1 5kb[1] ,细胞每经过一次分裂端粒缩短 50~ 2 0 0bp ,这种分子侵蚀作用使得细胞的分裂次数有了生理限制 ,从而限制了体细胞的寿命。一种逃避这种限制的机制是端粒酶的激活 ,因为端粒酶能弥补端粒的缩短 ,因此端粒酶被认为与细胞的永生化、肿瘤发生和细胞衰老密切相关。近来 ,组成人端粒酶复合物的 3个主要成分已被鉴定。人端粒酶RNA成分 (hTR)提… 相似文献
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Chiou SM Chiu CH Yang ST Yang JS Huang HY Kuo CL Chen PY Chung JG 《Neurochemical research》2012,37(8):1790-1800
This research focused on the induction of cytotoxic effects by danthron, a natural anthraquinone derivative on C6 rat glioma cells through exploring the means of cell death and the effects on mitochondrial function. We found that danthron decreased the percentage of viable C6 cells and induced cell morphological changes in a dose-and time-dependent manner. The morphological and nuclei changes (DAPI staining) in C6 cells were observed using a contrast-microscope and fluorescence microscopy, respectively. The results suggest that cell death of C6 cells which are induced by danthron is closely related to apoptotic death. Danthron decreased the level of mitochondrial membrane potential (ΔΨ( m )), stimulated the release of cytochrome c from mitochondria to cytosol and promoted the levels of caspase-9 and caspase-3, or induced the release of AIF and Endo G from mitochondria. Based on both observations, we suggest that the danthron-provoked apoptotic death of C6 cells is mediated through the mitochondria-dependent pathway. Furthermore, our results also indicated that danthron triggered apoptosis through reactive oxygen species (ROS) production which were increased after 1 h exposure of danthron, which was reversed by the ROS scavenger N-acetyl-L: -cysteine (NAC). As a consequence, danthron-mediated cell death of C6 cells via ROS production, mitochondrial transmembrane potential collapse and releases of cytochrome c, AIF and Endo G. Taken together, danthron was demonstrated to be effective in killing C6 rat glioma cells via the ROS-promoted and mitochondria-dependent apoptotic pathways. 相似文献
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Du Xuemei Fang Ting Liu Yan Huang Liying Wang Xiaoli Zhang Jie Cui Yangbo Zang Maosen Wang Guoying Fu Junjie Liu Yunjun 《In vitro cellular & developmental biology. Plant》2020,56(2):159-168
In Vitro Cellular & Developmental Biology - Plant - Callus induction in plants is similar to pluripotent stem cell induction in animals and can incite global changes in gene expression.... 相似文献
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Ermakov A. M. Gritsyna Yu. V. Belova S. P. Nemirovskaya T. L. Shenkman B. S. Vikhlyantsev I. M. 《Human physiology》2021,47(4):391-397
Human Physiology - Changes in titin alternative splicing in the rat soleus after seven-day gravitational unloading (the hindlimb unloading model) were studied by long-fragment PCR and nanopore... 相似文献
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Differential screening of a cDNA library prepared from mRNA of the hippocampus of estrogen-stimulated ovariectomized female rats led to the identification of a single estrogen-induced clone. Analysis of the sequence identified this cDNA as the gene coding for subunit III of the enzyme cytochrome c oxidase. Cytochrome c oxidase subunit III mRNA levels significantly increased as early as 3 h following the administration of a single dose of hormone. This effect was visible in the hippocampus and in the hypothalamus, but not in the other brain areas examined. Because subunit III of the cytochrome c oxidase is of mitochondrial origin, the mechanism involved in the estrogenic effect is still unknown. The observation that the activity of cytochrome c oxidase can also be induced by estrogens in the hippocampus indicates that this induction may be secondary to the increased expression of the other subunits of cytochrome c oxidase or to the general increase of neuronal activity. 相似文献
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Ping Liao Dejie Yu Zhenyu Hu Mui Cheng Liang Jue Jin Wang Chye Yun Yu Gandi Ng Tan Fong Yong Jia Lin Soon Yeow Leng Chua Tuck Wah Soong 《The Journal of biological chemistry》2015,290(14):9262-9272
L-type Cav1.2 Ca2+ channel undergoes extensive alternative splicing, generating functionally different channels. Alternatively spliced Cav1.2 Ca2+ channels have been found to be expressed in a tissue-specific manner or under pathological conditions. To provide a more comprehensive understanding of alternative splicing in Cav1.2 channel, we systematically investigated the splicing patterns in the neonatal and adult rat hearts. The neonatal heart expresses a novel 104-bp exon 33L at the IVS3-4 linker that is generated by the use of an alternative acceptor site. Inclusion of exon 33L causes frameshift and C-terminal truncation. Whole-cell electrophysiological recordings of Cav1.233L channels expressed in HEK 293 cells did not detect any current. However, when co-expressed with wild type Cav1.2 channels, Cav1.233L channels reduced the current density and altered the electrophysiological properties of the wild type Cav1.2 channels. Interestingly, the truncated 3.5-domain Cav1.233L channels also yielded a dominant negative effect on Cav1.3 channels, but not on Cav3.2 channels, suggesting that Cavβ subunits is required for Cav1.233L regulation. A biochemical study provided evidence that Cav1.233L channels enhanced protein degradation of wild type channels via the ubiquitin-proteasome system. Although the physiological significance of the Cav1.233L channels in neonatal heart is still unknown, our report demonstrates the ability of this novel truncated channel to modulate the activity of the functional Cav1.2 channels. Moreover, the human Cav1.2 channel also contains exon 33L that is developmentally regulated in heart. Unexpectedly, human exon 33L has a one-nucleotide insertion that allowed in-frame translation of a full Cav1.2 channel. An electrophysiological study showed that human Cav1.233L channel is a functional channel but conducts Ca2+ ions at a much lower level. 相似文献
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人端粒酶催化亚基基因启动子调控的肿瘤细胞特异表达载体 总被引:3,自引:0,他引:3
为获得端粒酶阳性肿瘤细胞特异表达载体用于癌症的基因治疗 ,克隆并构建了人端粒酶催化亚基 (hTERT)基因启动子调控的萤光素酶报告载体 .用脂质体转染法将其分别转染肿瘤细胞和正常细胞 ,检测其在肿瘤细胞和正常细胞中的转录活性 .hTERT启动子在所检测的 4种端粒酶阳性的肿瘤细胞中具有明显的转录活性 ,平均为阳性对照的 4 4 3% ;而在端粒酶阴性的正常人胚肺成纤维细胞中则无明显的转录活性 .提示hTRET启动子的转录活性在端粒酶阳性的肿瘤细胞中明显上调 ,由hTERT启动子构建的载体可能是一种新颖和有前景的肿瘤细胞特异性表达的基因治疗载体 相似文献
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Johannes Hradsky Hans-Gert Bernstein Monika Marunde Marina Mikhaylova Michael R. Kreutz 《The journal of histochemistry and cytochemistry》2015,63(10):793-804
Calneuron-1 and -2 are members of the neuronal calcium-binding protein family (nCaBP). They are transmembrane Calmodulin-like EF-hand Ca2+-sensors, and a function in the control of Golgi-to-plasma membrane vesicle trafficking has been assigned to both proteins. In this paper, we describe the distribution of Calneuron-1 in rat and human brains. We show that Calneuron-1 is ubiquitously expressed in all brain regions examined. The protein is most abundant in Purkinje cells of the cerebellum and principal neurons of the cortex and limbic brain whereas no expression in glial cells is apparent. In addition, we identify two novel splice isoforms of Calneuron-1 with extended N-termini. These isoforms are particular abundant in the cerebellum. Taken together, these data set grounds for a better understanding of the cellular function of Calneurons. 相似文献
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细胞凋亡(apoptosis)是多细胞生物的一种基本生命活动,在机体的生长发育、免疫调节及维持内环境稳定等各方面扮演着重要的角色.遗传和生化研究表明,细胞凋亡受到复杂而精细的调控.转录水平、翻译后水平等各种层次的调控,构成了一个复杂的凋亡调控网络. 相似文献