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Mechanism underlying the retarded nuclear translocation of androgen receptor splice variants

Science China(Life Sciences) 2019年第2期62卷 257-267页

作者： Ye Liu Yinyu Wang Fangfang Wang Jiexue Pan Jingjing Xu Jingyi Li Chengliang Zhou Guolian Ding Yanting Wu Xinmei Liu Jianzhong Sheng Hefeng Huang The International Peace Maternity and Child Health Hospital Shanghai Jiao Tong University School of Medicine The Key Laboratory of Reproductive Genetics (Ministry of Education) Zhejiang University Department of Pathophysiology Zhejiang University School of Medicine

As shown in our previous study, two alternatively spliced androgen receptor(AR) variants, which are exclusively expressed in the granulosa cells of patients with polycystic ovary syndrome, exhibit retarded nuclear translocation compared with wild-type AR. However, researchers have not yet determined whether these abnormalities correlate with heat shock protein 90(HSP90)and importin α(the former is a generally accepted co-chaperone of AR, and the latter is a component of classical nuclear import complexes). Here, these two variants were mainly retained in cytoplasm with HSP90 and importin α in the presence of dihydrotestosterone(DHT), and their levels in nucleus were significantly reduced, according to the immunofluorescence staining. The binding affinity of two AR variants for importin α was consistently decreased, while it was increased in WT-AR following DHT stimulation, leading to reduced nuclear import, particularly for the insertion-AR(Ins-AR). However, the binding affinities of two AR variants for HSP90 were increased in the absence of DHT compared with WT-AR, which functioned to maintain spatial structural stability, particularly for the deletion-AR(Del-AR). Therefore, the retarded nuclear translocation of two AR variants is associated with HSP90 and importin α, and the abnormal binding affinities for them play critical roles in this process.

关键词： androgen receptor splice variants nuclear translocation heat shock protein 90 importin α polycystic ovary syndrome

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Differential expression of glial cell line-derived neurotrophic factor splice variants in the mouse brain

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Neural Regeneration Research 2020年第2期15卷 270-276页

作者： Xiao-He Gu Heng Li Lin Zhang Tao He Xiang Chai He Wei Dian-Shuai Gao Department of Anatomy and Neurobiology Xuzhou Medical UniversityXuzhouJiangsu ProvinceChina Department of Neurosurgery First Affiliated Hospital of Soochow UniversitySuzhouJiangsu ProvinceChina

Glial cell line-derived neurotrophic factor(GDNF) plays a critical role in neuronal survival and function. GDNF has two major splice variants in the brain,α-pro-GDNF and β-pro-GDNF, and both isoforms have strong neuroprotective effects on dopamine neurons. However, the expression of the GDNF splice variants in dopaminergic neurons in the brain remains unclear. Therefore, in this study, we investigated the mRNA and protein expression of α-and β-pro-GDNF in the mouse brain by real-time quantitative polymerase chain reaction, using splice variant-specific primers, and western blot analysis. At the mRNA level,β-pro-GDNF expression was significantly greater than that of α-pro-GDNF in the mouse brain. In contrast, at the protein level,α-pro-GDNF expression was markedly greater than that of β-pro-GDNF. To clarify the mechanism underlying this inverse relationship in mRNA and protein expression levels of the GDNF splice variants, we analyzed the expression of sorting protein-related receptor with A-type repeats(SorLA) by real-time quantitative polymerase chain reaction. At the mRNA level, SorLA was positively associated with β-pro-GDNF expression, but not with α-pro-GDNF expression. This suggests that the differential expression of α-and β-pro-GDNF in the mouse brain is related to SorLA expression. As a sorting protein, SorLA could contribute to the inverse relationship among the mRNA and protein levels of the GDNF isoforms. This study was approved by the Animal Ethics Committee of Xuzhou Medical University, China on July 14, 2016.

关键词： Δ78 locus brain region dopaminergic neurons glial cell line-derived neurotrophic factor mouse brain precursor protein α-pro-GDNF β-pro-GDNF sorting protein-related receptor with A-type repeats splice variants

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A nucleotide substitution at the 5′ splice site of intron 1 of rice HEADING DATE 1(HD1) gene homolog in foxtail millet, broadly found in landraces from Europe and Asia

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The Crop Journal 2015年第6期3卷 481-488页

作者： Kenji Fukunaga Naoko Izuka Takehiro Hachiken Satoshi Mizuguchi Hidemi Ito Katsuyuki Ichitani Faculty of Life and Environmental Sciences Prefectural University of Hiroshima Faculty of Agriculture Kagoshima University

We investigated genetic variation of a rice HEADING DATE 1(HD1) homolog in foxtail millet.First, we searched for a rice HD1 homolog in a foxtail millet genome sequence and designed primers to amplify the entire coding sequence of the gene. We compared full HD1 gene sequences of 11 accessions(including Yugu 1, a Chinese cultivar used for genome sequencing) from various regions in Europe and Asia, found a nucleotide substitution at a putative splice site of intron 1, and designated the accessions with the nucleotide substitution as carrying a splicing variant. We verified by RT-PCR that this single nucleotide substitution causes aberrant splicing of intron 1. We investigated the geographical distribution of the splicing variant in 480 accessions of foxtail millet from various regions of Europe and Asia and part of Africa by d CAPS and found that the splicing variant is broadly distributed in Europe and Asia. Differences of heading times between accessions with wild type allele of the HD1 gene and those with the splicing variant allele were unclear. We also investigated variation in 13 accessions of ssp. viridis, the wild ancestor, and the results suggested that the wild type is predominant in the wild ancestor.

关键词： Foxtail millet Geographical distribution HD1(HEADING DATE 1) homolog Setaria italica splice site

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Correction of RNA aberrant splice increases foreign gene expression in transgenic mice

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Chinese Science Bulletin 1999年第3期44卷 221-225页

作者： Yifan Lu Chai Tian Jixian Deng Peitang Huang Acad Mil Med Sci Inst Biotechnique Beijing 100071 Peoples R China Chinese Acad Med Sci Cardiovasc Inst Beijing 100037 Peoples R China Chinese Acad Med Sci Fuwai Hosp Beijing 100037 Peoples R China

Transgenic mice with mammary gland secreting human granulocyte colony stimulating factor (G-CSF) were produced using mice whey acid protein gene promoter. It was found that there was very low expression level in mammary gland. Human G-CSF cDNA was obtained by RT-PCR from transgenic mice mammary gland. Sequence analysis showed that this G-CSF gene deleted the 4th exon, and compared with human G-CSF genomic DNA, there were donor and acceptor splice sites in the deletion fragment. It was considered that the 3rd and 4th introns also delete in G-CSF fragment. The transgenic construct was corrected by deleting the 3rd and 4th introns to construct the minigene, which was used to produce transgenic mice by microinjection. Northern blot showed that G-CSF expression using the new construct increased 5.4 times as that before in transgenic mice. The results suggested that it was possible that RNA aberrant splice result in low expression in transgenic mice.

关键词： transgenic RNA aberrant splice expression.

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Theoretical Analysis of the Factors Affecting splice Loss During Fabric Spreading

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Journal of China Textile University(English Edition) 2000年第2期17卷 65-68页

作者： Hui Chi Leung Patrick and NG Sau Fun FrencyInstitute of Textiles & Clothing, The Hong Kong Polytechnic University The Hong Kong Polytechnic Univ Hong Kong Hong Kong

Material utilization is mainly concerned by industrialpractitioners since it contributes to minimize the totalproduction costs for garment manufacturing. Apart fromthe marker loss, splice loss (the fabric loss during thejoining of different fabric rolls) is one of the primarycauses of fabric loss, but it has not been considered spe-cifically in the past literatures. The amount of splice lossduring fabric spreading is governed by various factors in-cluding the marker length, the number of fabric rolls,and the number of splicing intervals for laying-up. Inour study, these three effects on splice loss during fabricspreading were computed and analysed.

关键词： material utilization spreading splice loss

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CircAST:Full-length Assembly and Quantification of Alternatively spliced Isoforms in Circular RNAs

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Genomics, Proteomics & Bioinformatics 2019年第5期17卷 522-534页

作者： Jing Wu Yan Li Cheng Wang Yiqiang Cui Tianyi Xu Chang Wang Xiao Wang Jiahao Sha Bin Jiang Kai Wang Zhibin Hu Xuejiang Guo Xiaofeng Song Department of Biomedical Engineering Nanjing University of Aeronautics and AstronauticsNanjing 211106China School of Biomedical Engineering and Informatics Nanjing Medical UniversityNanjing 211166China State Key Laboratory of Reproductive Medicine Nanjing Medical UniversityNanjing 211166China Center of Pathology and Clinical Laboratory Sir Run Run HospitalNanjing Medical UniversityNanjing 211166China Center for Cellular and Molecular Therapeutic Children’s Hospital of PhiladelphiaPhiladelphiaPA 19104USA

Circular RNAs(circ RNAs),covalently closed continuous RNA loops,are generated from cognate linear RNAs through back splicing events,and alternative splicing events may generate different circ RNA isoforms at the same locus.However,the challenges of reconstruction and quantification of alternatively spliced full-length circ RNAs remain unresolved.On the basis of the internal structural characteristics of circ RNAs,we developed Circ AST,a tool to assemble alternatively spliced circ RNA transcripts and estimate their expression by using multiple splice graphs.Simulation studies showed that Circ AST correctly assembled the full sequences of circ RNAs with a sensitivity of 85.63%–94.32%and a precision of 81.96%–87.55%.By assigning reads to specific isoforms,Circ AST quantified the expression of circ RNA isoforms with correlation coefficients of 0.85–0.99 between theoretical and estimated values.We evaluated Circ AST on an in-house mouse testis RNA-seq dataset with RNase R treatment for enriching circ RNAs and identified 380 circ RNAs with full-length sequences different from those of their corresponding cognate linear RNAs.RT-PCR and Sanger sequencing analyses validated 32 out of 37 randomly selected isoforms,thus further indicating the good performance of Circ AST,especially for isoforms with low abundance.We also applied Circ AST to published experimental data and observed substantial diversity in circular transcripts across samples,thus suggesting that circ RNA expression is highly regulated.Circ AST can be accessed freely at https://***/xiaofengsong/CircAST.

关键词： Circular RNA Full-length reconstruction Isoform quantification Multiple splice graph model Transcriptome

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Impact of taxanes on androgen receptor signaling

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Asian Journal of Andrology 2019年第3期21卷 249-252页

作者： Shanshan Bai Bryan Y Zhang Yan Dong College of Life Sciences Jilin University Changchun 130012 China Department of Structural and Cellular Biology Tulane University School of Medicine Tulane Cancer Center New Orleans LA 70112 USA Lusher Charter School New Orleans LA 70115 USA

The development and progression of metastatic castration-resistant prostate cancer is the major challenge in the treatment of advanced prostate cancer. The androgen receptor signaling pathway remains active in metastatic castration-resistant prostate cancer. Docetaxel and cabazitaxel are the first- and second-line chemotherapy, respectively, for patients with metastatic castration-resistant prostate cancer. These two taxanes, in general, function by (i) inhibiting mitosis and inducing apoptosis and (ii) preventing microtubule-dependent cargo trafficking. In prostate cancer, taxanes have been reported to inhibit the nuclear translocation and activity of the androgen receptor. However, whether this is attainable or not clinically remains controversial. In this review, we will provide a comprehensive view of the effects of taxanes on androgen receptor signaling in prostate cancer.

关键词： androgen receptor androgen receptor splice variant cabazitaxel docetaxel prostate cancer taxane

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Experimental seismic behavior of squat shear walls with precast concrete hollow moulds

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Earthquake Engineering and Engineering Vibration 2019年第4期18卷 871-886页

作者： Han Wenlong Zhao Zuozhou Qian Jiaru Zhang Yingbao Ma Tao Key Laboratory of Civil Engineering Safety and Durability of China Education Ministry Tsinghua University Beijing 100084 China China Institute of Building Standard Design & Research Beijing 100048 China Beijing Everest Green Building Technology Ltd. Beijing 102209 China Beijing Institute of Architectural Design Beijing 100045 China

This study proposes an innovative precast shear wall system, called an EVE precast hollow shear wall structure (EVE-PHSW). Precast panels in EVE-PHSW are simultaneously precast with vertical and horizontal holes. Noncontact lap splices of rebars are used in vertical joints connecting adjacent precast panels for automated prefabrication and easy in situ erection. The seismic behavior of EVE walls was examined through a series of tests on six wall specimens with aspect ratios of 1.0~1.3. Test results showed that EVE wall specimens with inside cast-in situ concrete achieved the desired “strong bending and weak shear” and failed in shear mode. Common main diagonal cracks and brittle shear failure in squat cast-in situ walls were prevented. Inside cast-in situ concrete could signifi cantly improve the shear strength and stiff ness of EVE walls. The details of boundary elements (cast-in situ or prefabricated) and vertical joints (contiguous or spaced) had little eff ect on the global behavior of EVE walls. Noncontact lap splices in vertical joints could enable EVE walls to exhibit stable load-carrying capacity through extensive deformations. Evaluation on design codes revealed that both JGJ 3-2010 and ACI 318-14 provide conservative estimation of shear strength of EVE walls, and EVE walls achieved shear strength reserves comparative to cast-in situ walls. The recommended eff ective stiff ness for cast-in situ walls in ASCE 41-17 appeared to be appropriate for EVE walls.

关键词： precast concrete hollow mould low-aspect-ratio noncontact lap splice prefabricated boundary element vertical and horizontal joints seismic behavior

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Hepatitis B virus pre-S/S variants in liver diseases

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World Journal of Gastroenterology 2018年第14期24卷 1507-1520页

作者： Bing-Fang Chen School of Medicine Fu-Jen Catholic University

Chronic hepatitis B is a global health problem. The clinical outcomes of chronic hepatitis B infection include asymptomatic carrier state, chronic hepatitis(CH), liver cirrhosis(LC), and hepatocellular carcinoma(HCC). Because of the spontaneous error rate inherent to viral reverse transcriptase, the hepatitis B virus(HBV) genome evolves during the course of infection under the antiviral pressure of host immunity. The clinical significance of pre-S/S variants has become increasingly recognized in patients with chronic HBV infection. Pre-S/S variants are often identified in hepatitis B carriers with CH, LC, and HCC, which suggests that these naturally occurring pre-S/S variants may contribute to the development of progressive liver damage and hepatocarcinogenesis. This paper reviews the function of the pre-S/S region along with recent findings related to the role of pre-S/S variants in liver diseases. According to the mutation type, five pre-S/S variants have been identified: pre-S deletion, pre-S point mutation, pre-S1 splice variant, C-terminus S point mutation, and pre-S/S nonsense mutation. Their associations with HBV genotype and the possible pathogenesis of pre-S/S variants are discussed. Different pre-S/S variants cause liver diseases through different mechanisms. Most cause the intracellular retention of HBV envelope proteins and induction of endoplasmic reticulum stress, which results in liver diseases. Pre-S/S variants should be routinely determined in HBV carriers to help identify individuals who may be at a high risk of less favorable liver disease progression. Additional investigations are required to explore the molecular mechanisms of the pre-S/S variants involved in the pathogenesis of each stage of liver disease.

关键词： hepatitis B virus pre-S/S mutant pre-S deletion splice variant spPS1 chronic hepatitis liver cirrhosis hepatocellular carcinoma

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splice加速及并行SPICE模型构想及其在Transputer实现

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小型微型计算机系统 1990年第4期11卷 28-31页

作者：刘莘清华大学计算机系

在VLSI设计领域,CAE(Computer Aided Engincering)已成为一个基本的设计工具。在设计过程的诸个环节中,电路分析(又称电路模拟)是相当耗时的工作之一。有人做过估计:CAE工作站运算量的 2/3花在模拟上,对于某些大规模设计工作而言,主机... 详细信息

在VLSI设计领域,CAE(Computer Aided Engincering)已成为一个基本的设计工具。在设计过程的诸个环节中,电路分析(又称电路模拟)是相当耗时的工作之一。有人做过估计:CAE工作站运算量的 2/3花在模拟上,对于某些大规模设计工作而言,主机仍会遇到运算瓶颈,因此加速研究显得十分必要。本文介绍利用Transputer对混合模拟软件splice所做的加速工作,加速效果十分显著;此外,Transputer的特长还在于它的并行性,为此,本文也提出一种利用Transputer实现并行SPICE模型构想。

关键词： splice Transputer VLSI 集成电路

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