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王永樑

王永樑(Robert Y.-L. Wang)

職稱:副教授

研究室名稱:分子病毒研究室 (第一醫學大樓十樓B區)

最高學歷:Ph.D.

學校/國家:國立台灣大學微生物與生化學所

分機號碼:3691

電子郵件帳號yuwang@mail.cgu.edu.tw

個人網頁網址https://sites.google.com/site/cgu3691/

研究室現有:

博士後研究員0

博士班研究生2

碩士班研究生3

專任研究助理3

大學部專題生5

 

研究方向及研究室特色

實驗室研究主題:腸病毒 71 型及日本腦炎病毒之病毒複製和宿主之間的相互作用機制。

我們實驗室主要從事高深的病毒與寄主互相作用的研究。目前實驗室執行國科會計畫有兩大方向: () 嘗試將兩種正股 RNA 病毒 (腸病毒 71 Enterovirus 71EV71 日本腦炎, Japanese Encephalitis VirusJEV),轉型進入酵母菌中,測試其是否可以利用酵母菌為宿主進行複製;進一步利用這種最簡單的真核生物,進行病毒與寄主蛋白的互動機制研究。這項研究策略是基於過去我在美國肯塔基大學病理學部門進行研究工作。我使用tombusvirus(一種感染於植物的 positive stranded RNA病毒)作為一個模型。加上以 Yeast 為宿主進行實驗,成功地證明Glyceraldehyde-3-phosphate dehydrogenas(GAPDH),一種著名的代謝酵素,能透過它的選擇性結合不同股 RNA 的能力提升RNA 病毒的複製。 這重要發現將領導病毒學家理解病毒能利用寄主細胞內原有的蛋白質,提供本身複製和演化的一項工具。 這也提供研究對抗病毒及病毒傳染病作戰的一個新的策略及線索。我的這項研究工作也發表於 Cell Host&Microbe , 這是全世界最好的研究病毒及寄主互相作用雜誌之一

() 利用蛋白質體學方法,分析受到腸病毒 71 型及日本腦炎病毒感染後細胞之外分泌蛋白的組成,並且探討其可能影響病毒於體內複製,體外感染的機制。我的主要研究方向如下:

1. 利用系統生物學的方式,包括基因組及蛋白質體的篩選方式,尋找可能參與/影響 RNA 病毒複製的宿主蛋白。進而研究這些蛋白對於病毒複製的角色與實際參與的功能。

2. 分析及功能性闡述三種 RNA 結合性宿主蛋白於 RNA 病毒複製過程中參與的角色。包括:Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), p68 DEAD-box RNA helicase homolog Dp2 以及蛋白轉譯延長因子 (EF-1 alpha)

3. 負責液泡性蛋白分煉之功能性蛋白對於 RNA 病毒複製的角色。

 

The goals of Dr. Wang’s research are focused on the interaction of host factors and RNA viruses. Their first project is in the field of developing of yeast as a model host and powerful in vitro assay with purified replicase complexes, together with reverse genetic approaches that make Enterovirus 71 (EV71), Japanese Encephalitis Virus, capable of replicating in yeast, as to study fundamental aspects of viral RNA replication. The second project, Dr. Wang’s laboratory will use proteomic approaches to analyze infection of two cell lineages, RD cells and SF268 cells, with EV71, focusing on the secreted proteins. First, they will identify several secreted proteins which are found only in the EV71-infected cells by the combination using of one dimension electrophoresis and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Second, they will perform 2D-electrophoresis followed by MALDI-TOF/TOF in order to validate those proteins. Once they identify the host proteins which were presented in the secreted proteins of EV71-infected cells, they will discuss the results obtained taking into account the proteins previously described in the secretome of virus-infected mammalian cells, proteins present in human plasma and proteins of interest for EV71 pathogenesis. Altogether these data will provide clues for the progress in the understanding of the role of host secretion in disease progression.The specific aims are:

1) Identification of numerous host factors affecting/involving viral replication by using a systems biology approach, including genome-wide screens and proteomics approaches.

2) Dissecting the roles of three RNA-binding host proteins: glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the p68 DEAD-box RNA helicase homolog Dpb2 and translation elongation factor EF-1alpha (Tef1/2p), all of which bind to TBSV RNA and also known to interact with Hepatitis C virus, West Nile virus, dengue 4 virus and hepatitis A virus RNAs, suggesting that host factors might play similar roles in replication of these viruses.

3) Roles of Vacuolar Protein sorting proteins in RNA virus replication.

 

 

發表論文:

  1. Prabowo BA, Wang RY* (co-first author), Secario MK, Ou PT, Alom A, Liu JJ, Liu KC (2017) Rapid detection and quantification of Enterovirus 71 by a portable surface plasmon resonance biosensor. Biosens. Bioelectron. DOI: 10.1016/j.bios.2017.01.043 (SCI, impact: 7.476) * Equally contributed authors
  2. Lin JJ, Wang RY, Chen CJ, Chiu CC, Liao MH, Wu YJ (2016) Cytotoxicity of 11-epi-Sinulariolide Acetate Isolated from Cultured Soft Corals on HA22T Cells through the Endoplasmic Reticulum Stress Pathway and Mitochondrial Dysfunction. Int. J. Mol. Sci; 17, 1787/doi:10.3390 (SCI, impact factor: 3.257).
  3. Wang RY, Li YJ, Lee WC, Wu HH, Lin CY, Lee CC, Chen YC, Hung CC, Yang CW, Tian YC (2016) The association between polyomavirus BK strains and BKV viruria in liver transplant recipients. Scientific Reports; 6:28491 | DOI: 10.1038 (SCI, impact factor: 5.228)
  4. Wang RY, Weng KF, Huang YC, Chen CJ (2016) Elevated expression of circulating miR876-5p is a specific response to severe EV71 infection. Scientific Reports; 6:24149 | DOI: 10.1038 (SCI, impact factor: 5.228).
  5. Wang RY, Wu YJ, Chen HS, Chen CJ (2016) A KDEL retrieval system for ER-Golgi transport of Japanese Encephalitis viral particles. Viruses; 8 (44); doi: 10.3390/v8020044 (SCI, impact factor: 3.042).
  6. Wang RY, Kuo RL, Yen SM, Chu CH, Wu JY, Huang YC, Chen CJ (2015). Proteome demonstration of alpha-1-acid glycoprotein and alpha-1-antichymotrypsin are candidate biomarkers for diagnosis of Enterovirus 71 infection. The Pediatric Infectious Disease Journal; 34:304-310 (SCI, impact factor: 2.587).
  7. Kuo RL, Lin YH, Wang RY, Hsu CW, Chiu YT, Huang HI, Kao LT, Yu JS, Shih SR, Wu CC. (2015). Proteomics Analysis of EV71-Infected Cells Reveals the Involvement of Host Protein NEDD4L in EV71Replication. J. Proteome Res; 14:1818-1830. (SCI, impact factor: 4.173)
  8. Weng KF, Hung CT, Hsieh PT, Li ML, Chen GW, Kung YA, Huang PN, Kuo RL, Chen LL, Lin JY, Wang RY, Chen SJ, Tang P, Horng JT, Huang HI, Wang, JR, Ojcius DM, Brewer G, Shih SR (2014) A cytoplasmic RNA virus generates functional viral small RNAs and regulates viral IRES activity in mammalian cells. Nucleic Acids Res. 42:12789-12805. (SCI, impact factor: 9.202)
  9. Su C-C, Yang Z-Y, Su J-H, Din Z-H, Wang RY*(corresponding author), Wu Y-J* (2014) 13-Acetoxysarcocrassolide induces apoptosis on human gastric carcinoma cells through mitochondria-related apoptotic pathways: p38/JNK activation and PI3K/AKT suppression. Mar. Drugs; 12: 5295-5315. (SCI, impact factor: 3.345). * Equally contributed authors
  10. Liu Y-C, Kuo R-L, Lin J-Y, Huang P-N, Huang Y, Liu H, Arnold J, Chen S-J, Wang RY, Cameron C, Shih S-R (2014) Cytoplasmic Viral RNA-Dependent RNA Polymerase Disrupts the Intracellular Splicing Machinery by Entering the Nucleus and Interfering with Prp8. PLOS Pathog 10(6): e1004199. doi:10.1371/journal.ppat.1004199. (SCI, impact factor: 7.003)
  11. Huang HI, Chen SK, Wang RY, Shen CR, Cheng YC (2013) Human Foreskin-derived Fibroblast-like Stromal Cells are able to Differentiate into Functional Hepatocyte-like Cells. Cell Biol International; 37(12): 1308-1319. (SCI, impact factor: 1.663)
  12. Wang RY, Kuo RL, Ma WC, Huang HI, Yu JS, Yen SM, Huang CR, Shih SR (2013) Heat Shock Protein-90-beta Facilitates Enterovirus 71 Viral Particles Assembly. Virology; 443: 236-247 (SCI, impact factor: 3.200).
  13. Kuo RL, Kao Li-Ting, Lin SJ, Wang RY, Shih SR (2013) MDA5 plays a crucial role in Enterovirus 71 RNA mediated IRF3 activation. PLoS ONE 8(5): e63431. doi:10.1371/journal.pone.0063431(SCI, impact factor: 3.057)
  14. Neoh CA*, Wang RY* (co-first author), Din ZH, Su JH, Chen YK, Tsai FJ, Weng SH, Wu YJ (2012) Induction of Apoptosis by Sinulariolide from Soft Coral through Mitochondrial-Related and p38MAPK Pathways on Human Bladder Carcinoma Cells. Mar. Drugs; 10: 2893-2911. (SCI, impact factor: 3.345) * Equally contributed authors
  15. Huang CR, Wang RY, Hsu SC, Lo SJ (2012) Lysine-71 in the Large Delta Antigen of Hepatitis Delta Virus Clade 3 Modulates its Localization and Secretion. Virus Res; 170: 75-84. (SCI, impact factor: 2.526)
  16. Liu CI*, Wang RY* (co-first author), Lin JJ, Su JH, Chiu CC, Chen JC, Chen JY, Wu YJ (2012) Proteomic Profiling of the 11-Dehydrosinulariolide-treated Oral Carcinoma Cells Ca9-22: effects on the cell apoptosis through Mitochondrial-related and ER stress pathway. J. Proteomics; 75:5578-5589. (SCI, impact factor: 3.867) * Equally contributed authors
  17. Wang RY, Kui Li. (2012) Host factors in the replication of positive-strand RNA viruses. Chang Gung Med J; 35: 111-124.
  18. Li YJ, Wu HH, Weng CH, Chen YC, Hung CC, Yang CW, Wang RY, Sakamoto N, Tian YC. (2012) Cyclophilin A and nuclear factor of activated T cells are essential in cyclosporin A-mediated suppression of polyomavirus BK replication. American J Transplantation; doi:10.1111/j.1600-6143.2012.04116.x. (SCI, impact factor: 5.669)
  19. Hung CY, Tsai MC, Wu YP, Wang RY*. (2011)  Identification of heat shock protein 90 beta in JEV-induced secretion proteins. Journal of General Virology; 92:2803-2809. (SCI, impact factor: 3.192) *Corresponding author
  20. Prasanth KR, Huang YW, Liou MR, Wang RY, Hu CC, Tsai CH, Meng M, Lin NS, Hsu YH. (2011)  Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) negatively regulates the replication of Bamboo mosaic virus and its associated satellite RNA. Journal of Virology; 85: 8829-8840. (SCI, impact factor: 4.606)
  21. Nagy PD, Wang RY, Pogany J, Hafren A, Makinen K.  (2011) Emerging picture of host chaperone and cyclophilin roles in RNA virus replication. Virology; 411:374-382. (SCI, impact factor: 3.200)
  22. Wang RY, Huang YR,  Chong KM, Hung CY, Ke ZL, Chang RY. (2011)  DnaJ homolog Hdj2 Facilitates Japanese Encephalitis Virus Replication. Virology J; 8:471. (SCI, impact factor: 2.362)
  23. Wu YP, Chang CM, Schuyler S, Hung CY, Tsai MC, Wang RY*. (2011) Japanese Encephalitis Virus Co-opts the ER-Stress Response Protein GRP78 for Viral Infectivity. Virology J; 8:128. (SCI, impact factor: 2.362) *Corresponding author.
  24. Wang RY, Stork J, Pogany J, Nagy PD. (2009) A temperature sensitive mutant of heat shock protein 70 reveals an essential role during the early steps of tombusvirus replication. Virology; 394:28-38. (SCI, impact factor: 3.200).
  25. Wang RY, Stork J, Nagy PD (2009). A key role for heat shock protein 70 in localization and insertion of the tombusvirus replication proteins to intracellular membranes. J. Virology; 83:3276-3287 (SCI, impact factor: 4.606).
  26. Wang RY, Nagy PD (2008). Tomato bushy stunt virus Co-Opts the RNA-Binding Function of a Host Metabolic Enzyme for Viral Genomic RNA Synthesis. Cell Host&Microbe; 3:178-187. (SCI, impact factor: 12.552).
  27. Wang RY, Shen CN, Tosh D, Lin MH, Shih C (2005). Hepatocyte-like cells transdifferentiated from pancreatic origin can support replication of hepatitis B virus. J. Virology;79:13116-13128. (SCI, impact factor: 4.606).
  28. Chua PK, Wang RY, Suk FM, Lin MH, Masuda T, and Shih C (2005). A Naturally Occurring Mutation (L77R) of Hepatitis B Virus Small S Envelope Gene Results in Low-Level Virion Secretion and Can be Suppressed by Another Natural Envelope Mutation (W74L). J. Virology;79:13483-13496. (SCI, impact factor: 4.606).
  29. Wang YL, Wu CY, Chang CT, Sung HY (2003). Invertase inhibitors from sweet potato (Ipomoea Batatas): purification and biochemical characterization. J. Agric. Food Chem;51:4804-4809. (SCI, impact factor: 2.857)
  30. Wang YL, Lee YH, Chen CC, Chang CT, Sung HY (1998). Purification and characterization of soluble acid invertase from yam tubers. J. Chinese Agric. Chem. Soc;36: 464-472.
  31. Fu RH*, Wang YL* ,Sung HY (2002). Biochemical and molecular biological studies of plant invertases. Food Sci. Agric. Chem.;4:1-7 (*Equally  contribution).
  32. Fu RH, Wang YL, Sung HY. (2003) Cloning, characterization and functional expression of a new beta-D-fructofuranosidase (Osbetafruct2) cDNA from Oryza sativa. Biotechnol Lett. 25(6):455-459.

 

Conference Presentations:

 

  1. Wang RY, Ou PT (2016) Staufen proteins: multitalented entertainers in Enterovirus 71 infection cycle. HFMD International conference 2016, Biopolis, Signapore (Invited Speaker).
  2. Wang RY, Lee HY, Tian YC (2016) Characterization of the dominant noncoding control region of polyomavirus BK emerged during prolonged propagation. 2016 American Society for Virology 35th annual meeting, Virginia Tech University, VA, USA (Oral presentation)
  3. Wang RY, Chi CT (2015) Staufen protein modulates the enterovirus 71 infection cycle through its binding to viral RNA. 2015 American Society for Virology 34th annual meeting, Western University, ON, Canada (Oral presentation).
  4. Wang RY, Lee HY, Tian YC (2015) Characterization of the dominant noncoding control region of polyomavirus BK emerged during prolonged propagation. Department of Biological Sciences, Purdue University, IN, USA (Invited Speaker)
  5. Wang RY, Chen HS (2014) Profiling of small RNAs in JEV-infected BHK-21 cells. 2014 American Society for Virology 33rd annual meeting, Colorado State University, Fort Collins, CO, USA (Oral presentation).
  6. Wang RY, YX Li (2013) The role of KDEL motif of GRP78 in the Japanese Encephalitis Virus life cycle. 2013 American Society for Virology 32nd annual meeting, The Penn State University, PA, USA (Oral presentation).
  7. Wang RY (2012) Proteomic Profiling of the Enterovirus 71-induced Secretion Proteins reveals a role for Heat Shock Protein 90 beta in Facilitating Virus Particle Assembly. 2012 International symposium on RNA viruses, TaoYuan, TAIWAN (Invited speaker)
  8. Wang RY (2011) Detection of Hsp90 in the secretion medium from JEV-infected cells. 2011 International symposium on RNA viruses, TaoYuan, TAIWAN (Invited speaker)
  9. Wang RY, CY Hung, YP Wu, MC Tsai, S Schuyler (2011) The Detection of Heat Shock Protein 90 in the Secreted Proteins from Japanese Encephalitis Virus-Infected Cells. 2011 American Society for Virology 30th annual meeting, Minneapolis, MN, USA (Oral presentation).
  10. Wang RY (2010) Identification of the JEV-induced secretory proteins. 2010 International Symposium on RNA viruses, TaoYuan, TAIWAN. (Invited speaker)
  11. Wang RY, Nagy PD (2009) A key role for heat shock protein 70 in localization and insertion of the tombusvirus replication proteins to intracellular membrane. 2009 American Society for Virology 28th annual meeting, Vancouver, BC, Canada (Oral presentation).
  12. Wang RY, Nagy PD. (2007) A new role of a metabolic-RNA binding protein in tombusvirus RNA replication. 2007 American Society for Virology 26th annual meeting, Oregon, USA (Oral presentation).
  13. Wang RY, Shen CN, Tosh D, Lin MH, Shih C. (2004) Inducible replication of hepatitis B virus in transdifferentiated pancreatic hepatocytes. 2004 International hepatitis B virus meeting, Boston, USA (Oral presentation). 

Conference ABSTRACT:

 

  1. Lin CY, Wang RY* (2014) Hsa-miR-876-5p is down-regulated in the Enterovirus 71 persistent infected neuroblastoma cells The 29th Joint annual conference of Biomedical Science. (Poster Awarded).
  2. Huang CR, Yen SM, Chen HS, Chang YS, Wang RY* (2013) Profiling of small RNAs in JEV-infected BHK-21 cells. International Symposium on Infectious disease and signal transduction. National Cheng Kung University, Tainan, Taiwan.
  3. Lin YH, Wang RY (2011) Developing a Yeast system to study virus-host interaction in Enterovirus 71. 2011 International symposium on RNA viruses, TaoYuan, TAIWAN.
  4. Ma WC, Wang RY, Shih SR (2011) The role of heat shock protein 90 in Enterovirus 71 lifecycle. 2011 International symposium on RNA viruses, TaoYuan, TAIWAN.
  5. Chen TA, Chao M, Wang RY (2011) Replication of Hepatitis Delta Virus in yeast Saccharomyces Cerevisiae. 2011 International symposium on RNA viruses, TaoYuan, TAIWAN.
  6. Huang HH, Wang RY (2011) Identification of viral double-strand RNA specific binding proteins in JEV infected cells. 2011 International symposium on RNA viruses, TaoYuan, TAIWAN.
  7. SM Yen, Wang RY (2011) Investigation of Host Factors Involved in Replication of Defective RNA of Bamboo Mosaic Virus in Yeast. Nineteenth Symposium on Recent Advances in cellular and molecular biology, Kaoshiung, Taiwan.
  8. YP Wu, Wang RY (2011) Identification and characterization of molecular chaperone, GRP78, in Japanese Encephalitis Virus infected BHK21 cells. Nineteenth Symposium on Recent Advances in cellular and molecular biology, Kaoshiung, Taiwan.
  9. MC Tsai, Wang RY (2011) Characterization of the secretome in Japanese encephalitis virus-infected cells. Nineteenth Symposium on Recent Advances in cellular and molecular biology, Kaoshiung, Taiwan.
  10. CY Hung, Wang RY (2011) Detection of Heat shock protein 90 isoforms from JEV-infected cells. Nineteenth Symposium on Recent Advances in cellular and molecular biology, Kaoshiung, Taiwan.
  11. Wang RY, MC Tsai, CY Hung, TA Chen (2010) Identification of Japanese Encephalitis Virus-induced proteins in the cultured medium. 2010 American Society for Virology 29th annual meeting, Bozeman, MT, USA.
  12. Wang RY, MC Tsai (2010) Characterization of the secretory proteins in Japnaese Encephalitis Virus-infected cells. 9 th International symposium on Positive-strand RNA viruses, Atlanta, GA, USA.
  13. Wang RY, Tieu B, Shih C. (2004) Hepatitis B virus replication in a putative progenitor cell line with a differentiation potential to pancreas and liver. 2004 Pathology Department's 11th Annual Trainee Poster Session, University of Texas Medical Branch, Galveston, TX, USA.
  14. Wang RY, Sung HY. (2001) Invertases from sweet potato leaves: purificatioin, characterization and degradation. 17th Joint Annual Conference of Biochemical Sciences, Taipei, Taiwan

 

 

 

 

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