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Docking protein 6 (DOK6) selectively docks the neurotrophic signaling transduction to restrain peripheral neuropathy

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Signal Transduction and Targeted Therapy 2024年第3期9卷 1147-1164页

作者： Yan Guo Pan Xiang Xiaojiao Sun Wei Liu Jiafeng Zhou Bin Yin Lin Hou boqin qiang Huiliang Li Pengcheng Shu Xiaozhong Peng Department of Molecular Biology and Biochemistry Institute of Basic Medical SciencesMedical Primate Research CenterNeuroscience CenterChinese Academy of Medical SciencesSchool of Basic Medicine Peking Union Medical CollegeBeijingChina State Key Laboratory of Common Mechanism Research for Major Diseases Beijing 100005China Wolfson Institute for Biomedical Research University College LondonGower StreetLondon WC1E 6BTUK Chinese Institute for Brain Research Beijing 102206China State Key Laboratory of Respiratory Health and Multimorbidity Beijing 100005China Institute of Laboratory Animal Science Chinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100021China

The appropriate and specific response of nerve cells to various external cues is essential for the establishment and maintenance of neural circuits,and this process requires the proper recruitment of adaptor molecules to selectively activate downstream pathways.Here,we identified that DOK6,a member of the Dok(downstream of tyrosine kinases)family,is required for the maintenance of peripheral axons,and that loss of Dok6 can cause typical peripheral neuropathy symptoms in mice,manifested as impaired sensory,abnormal posture,paw deformities,blocked nerve conduction,and dysmyelination.Furthermore,Dok6 is highly expressed in peripheral neurons but not in Schwann cells,and genetic deletion of Dok6 in peripheral neurons led to typical peripheral myelin outfolding,axon destruction,and hindered retrograde axonal transport.Specifically,DOK6 acts as an adaptor protein for selectivitymediated neurotrophic signal transduction and retrograde transport for TrkC and Ret but not for TrkA and TrkB.DOK6 interacts with certain proteins in the trafficking machinery and controls their phosphorylation,including MAP1B,Tau and Dynein for axonal transport,and specifically activates the downstream ERK1/2 kinase pathway to maintain axonal survival and homeostasis.This finding provides new clues to potential insights into the pathogenesis and treatment of hereditary peripheral neuropathies and other degenerative diseases.

关键词： impaired restrain peripheral

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Polypyrimidine Tract-Binding Protein Regulates Enterovirus 71 Translation Through Interaction with the Internal Ribosomal Entry Site

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Virologica Sinica 2019年第1期34卷 66-77页

作者： Juemin Xi Fei Ye Guanzhou Wang Wei Han Zhizhong Wei Bin Yin Jiangang Yuan boqin qiang Xiaozhong Peng Institute of Medical Biology Chinese Academy of Medical Sciencesand Peking Union Medical College The State Key Laboratory of Medical Molecular Biology Department of Molecular Biology and BiochemistryInstitute of Basic Medical SciencesChinese Academy of Medical Sciences and Peking Union Medical College

Enterovirus 71(EV71), a major causative agent of hand, foot, and mouth disease, has caused periodic infection outbreaks in children in the Asia–Pacific region. In order to describe the largely unknown life cycle of EV71, the molecular basis of its virus-host interactions must first be determined. The 50 untranslated region of EV71 contains a cloverleaf-like structure and internal ribosomal entry site(IRES), which play an important role in transcription and translation of viral protein. We found that polypyrimidine tract-binding protein 1(PTB) bound to the IRES of EV71. RNA recognition motifs 1 and 2 of PTB were responsible for its binding to the EV71 IRES. Moreover, PTB protein was shuttled from nucleus to cytoplasm after EV71 infection. Additionally, IRES activity and viral protein production were inhibited by PTB knockdown. These results suggest that PTB interacts with the EV71 IRES, and positively regulates viral protein translation.

关键词： EV71 IRES PTB Translation

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Cloning of a novel gene encoding human thioredoxin-like protein

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Chinese Science Bulletin 1999年第18期44卷 1673-1676页

作者： Yan Zhou Meihui Pan Jiangang Yuan Guangwei Du boqin qiang Chinese Acad Med Sci Inst Basic Med Sci State Lab Med Mol Biol Beijing 100005 Peoples R China Peking Union Med Coll Beijing 100005 Peoples R China

mRNA differential display (DDRT-PCR) has been used to analyze different human fetal brain tissues of different developmental stages (13- and 33-week). According to the sequence of one EST obtained in this assay, a pair of primers have been designed to screen the arrayed human fetal brain cDNA library. A 1 .2-kb cDNA clone has been found. This cDNA con-

关键词： DDRT-PCR thioredoxin fetal brain.

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Cloning and identification of PHF2 cDNA and its alternatively spliced transcripts

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Chinese Science Bulletin 1999年第15期44卷 1382-1387页

作者： Junhua Wang Guangwei Du Yan Zhou Bin Yin Jiangang Yuan boqin qiang Chinese Acad Med Sci Inst Basic Med Sci State Key Lab Med Mol Biol New York NY 10005 USA

Genes encoding proteins with PHD (plant homeodomain) finger motif (C4HC3) are highly conserved from Arabidopsis to Homo sapiens. One of the major functions of these genes is regulating the expression of homeotic genes during the stage of embryonic development. They play a role in cell-cycle and cell differentiation and seem to be related with some human malignant diseases, such as leukemia. A human placenta cDNA library has been screened with cDNA probe amplified by PCR. The PGR primers have been designed according to the M96 (a mouse gene encoding a protein with PHD domain) homologous data in dbEST.A 2. 1 kb insert fragment in a positive cDNA clone has been isolated and sequenced. This new full-length cDNA is named PHF2 (GenBank Acc: AF052205). The putative protein composed of 567 aa has two typical PHD fingers at its N-terminus. Meanwhile it is identified that there are several alternatively spliced transcripts of PHF2 in different human tissues through the PCR amplification, Northern blot

关键词： PHD finger motif PHF2 cDNA alternative splicing.

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Isolation and expression pattern analysis of novel ESTs from human fetal brain

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Chinese Science Bulletin 1998年第21期43卷 1815-1819页

作者： Meihui Pan Jiangang Yuan Guangwei Du Yan Zhou Hui Yao boqin qiang Chinese Acad Med Sci Inst Basic Med Sci Natl Lab Med Mol Biol Beijing 100005 Peoples R China Paking Union Med Sci Beijing 100005 Peoples R China

In order to learn the mechanism of brain development and differentiation, 90 expressed sequence tags (ESTs) were isolated by differential display from the cerebrum and cerebellum of 13-week and 33-week fetal brains. After searching database, 74 of them represented novel genes, some of them were homologous to the brain development related genes. Using total cDNA probes, 79 of the ESTs and their expression differences in fetal brain were further characterized.

关键词： brain development and differentiation expressed sequence tags gene expression.

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Cloning and expression analysis of MBLL cDNA

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Chinese Science Bulletin 2000年第7期45卷 620-625页

作者： Guangwei Du Yan Zhou Jianhe Chen Junhua Wang Bin Yin Jiangang Yuan boqin qiang National Laboratory of Medical Molecular Biology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences and Peking Union Medical College National Center of Human Genome Beijing China

The mbl (musdeblind) gene of Drosophila encodes a nuclear protein which contains two Cys3His motifs. The mutation of mbl gene will disturb the differentiation of ali the Drosophila’s photoreceptors. Primers have been designed according to human EST086139, which is highly homologous to mbl gene. Human fetal brain cDNA library has been screened and a novel cDNA clone has been obtained. The 2595 bp cDNA, designated MBLL (muscleblind-like), contains an open reading frame which encodes 255 amino acids and has 4 Cys3His motifs (GenBank Acc. AF061261). The amino acids seguence shares high homology to Drosophila’s mbl. The Northern blot and RNA dot blot hybridization of 43 human adult tissues and 7 fetal tissues show that MBLL is a widely expressed gene, but the expression amounts differ in these tissues.

关键词： MSLL CyssHis zinc finger motif cDNA cloning tissue expression.

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Molecular dynamics simulation of site-directed mutagenesis of HIV-1 Tat trans-activator

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Chinese Science Bulletin 1999年第8期44卷 708-711页

作者： Yan Cui Lunjiang Ling Runsheng Chen Longchuan Bai Jiangang Yuan boqin qiang Chinese Acad Sci Inst Biophys Beijing 100101 Peoples R China Chinese Acad Med Sci Inst Basic Med Sci Beijing 100005 Peoples R China

The Cys-rich domain, core region and basic domain are highly conserved and very important to the trans-activation activity of HIV-1 Tat trans-activator. The three-dimensional structures of 6 mutants of HIV-1 Tat protein were constructed with the methods of molecular dynamics simulation. The variations of the structures of the mutants have been analyzed and the factors that led to abolishment of trans-activation activity have been discussed.

关键词： human immunodeficiency virus type 1 ( HIV-1) molecular dynamics simulation site-directed mutagenesis Tat protein.

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Cloning of human and mouse GRY-RBP cDNA

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Chinese Science Bulletin 2000年第4期45卷 343-350页

作者： Guangwei Du Meihui Pan Yan Zhou Jianhe Chen Hui Yao Jiangang Yuan boqin qiang Chinese Acad Med Sci Peking Union Med Coll Natl Ctr Human Genome Res Inst Inst Basic Med SciNatl Lab Med Mol Biol Beijing 100005 Peoples R China

After screening human fetal brain cDNA library, a novel human cDNA encoding an RNA-binding protein was cloned and named GRY-RBP. Human GRY-RBP cDNA is 2 932 bp in length, and encodes a protein of 623 amino acids with a deduced molecular mass of 69.6 ku. Human GRY-RBP contains three RNA recognition motifs (RRMs) and a glycine (G), tyrosin (Y)-rich C-terminus. Computer searches with sequence database revealed its moderate homology with RRMs of many RRM RNA-binding proteins. in this study, the full coding region and partial non-coding region of mouse GRY-RBP cDNA were amplified by PCR using mouse brain cDNA library as template. Three transcripts of Gry-rbp are expressed in ali tissues as shown by Northern blot, but the amounts in heart, planceta and skeletal muscle are considerably higher than those in other tissues.

关键词： RRM RNA-binding protein GRY-RBP tissue expression.

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RNA-binding Protein UNR Promotes Glioma Cell Migration and Regulates the Expression of Ribosomal Protein L9

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Chinese Medical Sciences Journal 2018年第3期33卷 143-151页

作者： Ningyu Tian Yingjiao Qi Yan Hu Bin Yin Jiangang Yuan boqin qiang Xiaozhong Peng Wei Han State Key Laboratory of Medical Molecular Biology Department of Molecular Biology and BiochemistryInstitute of Basic Medical Sciences Chinese Academy of Medical Sciences&School of Basic Medicine Peking Union Medical CollegeBeijing 100005China

Objective To investigate the role of RNA binding protein—upstream-of-N-Ras(UNR)in the development of glioma and its molecular mechanism.Methods First,bioinformatics analysis of CGGA database was performed to detect UNR expression level and prognosis of patients with glioma.Western blot and real-time PCR were used to detect UNR expression level in glioma cell lines and tissues.Next,UNR siRNAs were transfected in glioma cells,and MTS assay and scratch wound-healing assay were used to detect changes in cell proliferation and migration.Then,the candidate UNR target mRNAs were identified by analyzing the sequencing data of UNR iCLIP-seq,RNA sequencing and ribosome profiling databases of human melanoma.RNA immunoprecipitation and biotin pull-down assays were used to identify the UNR target mRNAs in glioma cells.Finally,western blot was used to detect the effect of UNR knockdown on ribosomal protein L9(RPL9)and RPL9 protein expression level in glioma cell lines.RPL9 siRNA was transfected in A172 and T98G and the expression of vimentin in the cells was detected with western blot.Results Bioinformatics analysis showed that UNR mRNA expression level was significantly higher in highgrade glioma[GradeⅡ(n=126),GradeⅢ(n=51),GradeⅣ(n=128),P<0.001].UNR high expression levels were associated with poor prognosis(P=0.0177).UNR had high expression level in glioma cell lines and patient samples compared with normal cell lines and normal brain samples(P<0.01).Knockdown of UNR inhibited glioma cells migration(P<0.05),but did not inhibit glioma cells growth in three glioma cell lines.UNR binded the 3’untranslated region(UTR)of PTEN and RPL9 mRNAs.RPL9 protein was significantly highly expressed in most glioma cell lines(n=9)and knockdown of UNR resulted in a downregulation of RPL9 protein expression.

关键词： UNR glioma migration ribosomal protein L9 vimentin

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Cloning and identification of a novel human glioma differentiation related gene GDR1

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Chinese Science Bulletin 2000年第11期45卷 995-999页

作者： Guangtao Li Hulin Jin Songnian Hu Jian Jin Chun Tu Jiangang Yuan boqin qiang Peking Union Med Coll Sch Basic MedNatl Ctr Human Genome Res Chinese Acad Med Sci Inst Basic Med SciNatl Lab Med Mol Biol Beijing 100005 Peoples R China

In order to identify the genes associated with glioblastoma differentiation, some ESTs, expressed differentially in the control cell and the differentiated human glioblastoma cell line BT-325 induced by the all-trans retinoid acid, have been isolated by the method of DDRT-PCR. Of the 46 ESTs sequenced, 19 are from new genes. A full-length 1 535-bp cDNA, termed gene GDR1, has been isolated from the human cDNA library using the probe designed according to one of the novel ESTs, HGBB098. The open reading frame of GDR1 gene encodes a putative protein containing 334 amino acid residues. Blast against the current GenBank DMA and protein sequence database did not reveal significant homology with any known proteins. RT-PCR shows that GDR1 mRNA level increased in the differentiated BT-325 cells after being treated with RA. The different expression patterns of GDR1 mRNA in human tissues have been detected through the multiple tissue Northern blot hybridization.

关键词： human glioma GDR1 RT-PCR.

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