郭德银
主要研究领域:RNA病毒(冠状病毒和丙肝病毒)的基因组复制和与宿主细胞相互作用的分子机理;病毒免疫逃逸的分子机制;基于RNA干扰(RNAi)的艾滋病毒基因治疗。
个性化签名
- 姓名:郭德银
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学术头衔:
博士生导师
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学科领域:
微生物学
- 研究兴趣:主要研究领域:RNA病毒(冠状病毒和丙肝病毒)的基因组复制和与宿主细胞相互作用的分子机理;病毒免疫逃逸的分子机制;基于RNA干扰(RNAi)的艾滋病毒基因治疗。
郭德银,男,1965年4月于河南省,博士毕业于联邦德国布伦瑞克理工大学,1996.1 - 2002.9,在芬兰赫尔辛基大学生物技术所做博士后和docent,现任武汉大学生命科学学院特聘教授,博导,副院长。
主要研究领域:RNA病毒(冠状病毒和丙肝病毒)的基因组复制和与宿主细胞相互作用的分子机理;病毒免疫逃逸的分子机制;基于RNA干扰(RNAi)的艾滋病毒基因治疗。
首次系统研究同一种Poty病毒所有编码蛋白和基因组RNA的相互关系,第一个建立起Potyvirus不同蛋白的相互作用关系图;筛选出两种与RNA静默病毒抑制蛋白相互作用的宿主蛋白质,为揭示RNA静默及其抗病毒的分子机理奠定了基础。2002年回国后主要从事肝炎病毒和艾滋病毒的研究工作,在SARS爆发后担任武汉大学SARS病毒攻关组组长,承担了国家重大基础研究"973"项目、国家自然科学基金等项目,在SARS病毒分离鉴定和基因组复制表达等方面做出了重要成绩,研究成果已经发表在国际最著名病毒学杂志《JVI》等期刊上。在使用RNA干扰技术进行艾滋病毒基因治疗方面也获得重要进展,结果已经发表在著名期刊《FEBS Lett》上。另外,本人协助著名病毒学家田波院士管理现代病毒学研究中心,迅速使该中心成为具有较大影响的病毒学研究基地;2003年由本人主持申请《教育部病毒学重点实验室》获得批准,为学科建设做出了贡献。
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成果数
13
郭德银, Snawar Hussain, † Ji'an Pan, † Yu Chen, Yalin Yang, Jing Xu, Yu Peng, Ying Wu, Zhaoyang Li, Ying Zhu, Po Tien, and Deyin Guo*
JOURNAL OF VIROLOGY, May 2005, p. 5288-5295,-0001,():
-1年11月30日
The expression of the genomic information of severe acute respiratory syndrome coronavirus (SARS CoV) involves synthesis of a nested set of subgenomic RNAs (sgRNAs) by discontinuous transcription. In ARS CoV-infected cells, 10 sgRNAs, including 2 novel ones, were identified, which were predicted to be functional in the expression of 12 open reading frames located in the 3 one-third of the genome. Surprisingly, one new sgRNA could lead to production of a truncated spike protein. Sequence analysis of the leader-body fusion sites of each sgRNA showed that the junction sequences and the orresponding transcription-regulatory sequence (TRS) are unique for each species of sgRNA and are consistent after virus passages. For the two novel sgRNAs, each used a variant of the TRS that has one nucleotide mismatch in the conserved hexanucleotide core (ACGAAC) in the TRS. Coexistence of both plus and minus strands of SARS CoV sgRNAs and evidence for derivation of the sgRNA core sequence from the body core sequence favor the model of discontinuous transcription during minus-strand synthesis. Moreover, one rare species of sgRNA has the junction sequence AAA, indicating that its transcription could result from a noncanonical transcription signal. Taken together, these results provide more insight into the molecular mechanisms of genome expression and subgenomic transcription of SARS CoV.
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郭德银, Zhaoyang Li, Yong Xiong, Yu Peng, Jian Pan, Yu Chen, Xiaoyun Wu, Snawar Hussain, Po Tien, Deyin Guo*
FEBS Letters 579(2005)3100-3106,-0001,():
-1年11月30日
RNA interference (RNAi), a sequence-specific RNA degradation mechanism mediated by small interfering RNA (siRNA), can be used not only as a research tool but also as a therapeutic strategy for viral infection. We demonstrated that intracellular expression of short hairpin RNA (shRNA) targeting human cyclin T1 (hCycT1), a cellular factor essential for transcription of messenger and genomic RNAs from the long terminal repeat promoter of provirus of human immunodeficiency virus type 1 (HIV-1), could effectively suppress the replication of HIV-1. We also showed that downregulation of hCycT1 did not cause apoptotic cell death, therefore, targeting cellular factor hCycT1 by shRNAs may provide an attractive approach for genetic therapy of HIV-1 infection in the future.
RNA interference, Human immunodeficiency virus type 1, Short hairpin RNA, Cyclin T1
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【期刊论文】Bovine PrPC directly interacts with aB-crystalline
郭德银, Guihong Sun a, b, Mingxiong Guo a, Ao Shen a, Fanghua Mei a, Xiaoxue Peng a, Rui Gong a, Deyin Guo a, Jianguo Wu a, Po Tien a, Gengfu Xiao a, *
FEBS Letters 579(2005)5419-5424,-0001,():
-1年11月30日
We used a bovine brain cDNA library to perform a yeast two-hybrid assay with bovine mature PrPC as bait. The screening result showed that aB-crystalline interacted with PrPC. The interaction was further evaluated both in vivo and in vitro with different methods, such as immunofluorescent colocalization, native polyacrylamide-gel electrophoresis, and IAsys biosensor assays. The results suggested that aB-crystalline may have the ability to refold denatured prion proteins, and provided first evidence that aB-crystalline is directly associated with prion protein.
Protein interaction, Prion, PrPC, aB-Crystalline, Small heat-shock protein
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郭德银, Deyin Guo, , Carl Spetz, Mart Saarma, and Jari P. T. Valkonen
MPMI Vol. 16, No.5, 2003, pp. 405-410.,-0001,():
-1年11月30日
Potyviral helper-component proteinase (HCpro) is a multifunctional protein exerting its cellular functions in interaction with putative host proteins. In this study, cellular protein partners of the HCpro encoded by Potato virus A (PVA)(genus Potyvirus) were screened in a potato leaf cDNA library using a yeast two-hybrid system. Two cellular proteins were obtained that interact specifically with PVA HCpro in yeast and in the two in vitro binding assays used. Both proteins are encoded by single-copy genes in the potato genome. Analysis of the deduced amino acid sequences revealed that one (HIP1) of the two HCpro interactors is a novel RING finger protein. The sequence of the other protein (HIP2) showed no resemblance to the protein sequences available from databanks and has known biological functions.
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【期刊论文】Hantavirus nucleocapsid protein interacts with the Fas-mediated apoptosis enhancer Daxx
郭德银, Xiao-Dong Li, Tomi P. Ma kela, Deyin Guo, Rabah Soliymani, Vesa Koistinen, Olli Vapalahti, Antti Vaheri and Hilkka Lankinen
Journal of General Virology (2002), 83, 759-766.,-0001,():
-1年11月30日
Hantaviruses cause two severe diseases, haemorrhagic fever with renal syndrome in Eurasia and hantavirus pulmonary syndrome in the Americas. To understand more about the molecular mechanisms that lead to these diseases, the associations of Puumala virus nucleocapsid protein (PUUV-N) with cellular proteins were studied by yeast two-hybrid screening. Daxx, known as an apoptosis enhancer, was identified from a HeLa cDNA library and its interaction with PUUV-N was confirmed by GST pull-down assay, co-immunoprecipitation and co-localization studies. Furthermore, domains of interaction were mapped to the carboxyl-terminal region of 142 amino acids in Daxx and the carboxyl-terminal 57 residues in PUUV-N, respectively. In pepscan assays, the binding sites of Daxx to PUUV-N were mapped further to two lysine-rich regions, of which one overlaps the sequence of the predicted nuclear localization signal of Daxx. These data suggest a direct link between host cell machinery and a hantavirus structural component.
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郭德银, Deyin Guo, Minna-Liisa Rajama ki, , Mart Saarma and Jari P. T. Valkonen
Journal of General Virology (2001), 82, 935-939.,-0001,():
-1年11月30日
A map for the interactions of the major proteins from Potato virus A (PVA) and Pea seed-borne mosaic virus (PSbMV) (members of the genus Potyvirus, family Potyviridae) was generated using the yeast two-hybrid system (YTHS). Interactions were readily detected with five PVA protein combinations (HC-HC, HC-CI, VPg-VPg, NIa-NIb and CP-CP) and weak but reproducible interactions were detected for seven additional combinations (P1-CI, P3-NIb, NIaPro-NIb, VPg-NIa, VPg-NIaPro, NIaPro-NIa and NIa-NIa). In PSbMV, readily detectable interactions were found in five protein combinations (HC-HC, VPg-VPg, VPg-NIa, NIa-NIa and NIa-NIb) and weaker but reproducible interactions were detected for three additional combinations (P3-NIa, NIa-NIaPro and CP-CP). The self-interactions of HC, VPg, NIa and CP and the interactions of VPg-NIa, NIa-NIaPro and NIa-NIb were, therefore, common for the two potyviruses. The multiple protein interactions revealed in this study shed light on the co-ordinated functions of potyviral proteins involved in virus movement and replication.
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郭德银, Igor Oruetxebarria a, Deyin Guo b, Andres Merits b, Kristiina Makinen b, Mart Saarma b, Jari P. T. Valkonen a, *
Virus Research 73(2001)103-112,-0001,():
-1年11月30日
Viruses of the genus Poty6irus, the largest genus of plant-infecting viruses, have a messenger-polarity ssRNA genome encapsidated by approximately 2000 units of the viral coat protein (CP), resulting in filamentous virions. Only few studies have examined potyvirus virions for the presence of other structural proteins. A protein linked covalently to the 5%-end of the genome has been identified in Tobacco 6ein mottling 6irus (TVMV) and Tobacco etch 6irus (TEV). In TEV, it is either the viral NIa protein or only its N-terminal domain (VPg) separated autocatalytically from the C-terminal proteinase domain (NIa-Pro). Virions of TVMV carry only the VPg. We examined virions of Potato 6irus A (PVA) for the genome-linked protein using immunoblotting or iodination and immunoprecipitation. The VPg (25kDa) only, and not the unprocessed NIa, was detected. Another signal corresponding to 49 kDa was detected in disrupted, RNase-treated virions with anti-VPg antibodies but not with antibodies to NIa-Pro. Since it possibly represented a dimeric form of the VPg, self-interaction of the VPg was tested using the yeast two-hybrid system, which showed that the VPg self-interacts in the absence of viral RNA.
Potato virus A, Poty6irus, Tobacco vein mottling virus
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郭德银, Andres Merits, , Oleg N. Fedorkin, Deyin Guo, Natalia O. Kalinina and Sergey Yu. Morozov
Journal of General Virology (2000), 81, 3099-3106.,-0001,():
-1年11月30日
The putative replication initiation protein (Rep) of Coconut foliar decay virus (CFDV) was expressed as a 6¬ His recombinant protein in E. coli and in recombinant baculovirus. Purified 6¬ His-Rep protein was demonstrated to possess sequence non-specific RNA- and ssDNA-binding activities as well as magnesium-dependent ATPase/GTPase activity. The yeast two-hybrid system revealed that CFDV Rep could interact with itself. Subcellular distribution of the CFDV Rep was studied by fractionation of insect cells infected with recombinant baculovirus expressing the 6¬ His–Rep protein and by laser scanning confocal microscopy of Nicotiana benthamiana epidermal cells bombarded with a construct encoding CFDV Rep fused to GFP. It was shown that CFDV Rep associated predominantly with nuclei and membranes of infected/transfected cells. These activities of CFDV-encoded Rep are very similar to those reported for Reps of geminiviruses.
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郭德银, Pa
Mechanisms of Development 93(2000)169-173,-0001,():
-1年11月30日
Dysfunction and downregulation of dad (defending against death) has been linked to programmed cell death (PCD) in animals and plants. As DAD is an essential subunit of the oligosaccharyltransferase that is located in the ER membrane, the results have raised the possibility that downregulation of N-linked glycosylation could be involved in the regulation of PCD. Here we show that the 16 kDa subunit of phytepsin, a vacuolar proteinase, is normally processed and glycosylated at the onset of DNA fragmentation in germinating barley scutella. Two cDNA clones encoding dad (dad1, dad2), and one cDNA encoding another subunit of the same oligosaccharyltransferase complex (ost1) were isolated from barley. Northern analysis of germinating scutella show that the expression of only dad1 is declining before onset of DNA fragmentation. In contrast to this, the expression of both dad2 and ost1 increase before onset of DNA fragmentation. q2000 Elsevier Science Ireland Ltd. All rights reserved.
N-linked glycosylation, Programmed cell death, Phytepsin, dad1, dad2, Ribophorin I, ost1
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郭德银, Deyin Guo, Andres Merits and Mart Saarma
Journal of General Virology (1999), 80, 1127-1131,-0001,():
-1年11月30日
Potyviral helper component-proteinase (HC-Pro) is a multifunctional protein involved in aphid transmission, long-distance movement, polyprotein processing, genome amplification and symptom expression. It has been proposed that the active form of HC-Pro is a dimer and that coat protein (CP) -HCPro interaction is required for aphid transmission. To test these proposed interactions between CP and HC-Pro of potato A potyvirus (PVA), the yeast two-hybrid system was used. HC-Pro was shown to interact with itself in vivo in yeast cells, as did CP. Taken together with previous observations, we conclude that the functional HC-Pro is a homodimer. Deletion analysis showed that a 24 aa domain in the N-terminal half and the C-terminal proteinase part of HC-Pro were required for the interaction between HC-Pro molecules. No interactions were found between HC-Pro and CP using the genes of aphid-transmissible as well as aphid non-transmissible strains of PVA.
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