SHIN-LIAN DOONG (董馨蓮)ORCID Follow
|Profile 2016-07-28 15:06:01|
|Education||Ph.D. Human Oncology University of Wisconsin at Madison.|
|Career and Experience||
Present : Associate Professor, Graduate Institute of Microbiology, College of Medicine, National Taiwan University.
1. Visiting Associate Professor, Graduate Institute of Microbiology, College of Medicine, National Taiwan University (1991.10-1993.07)
2. Postdoc. Research Associate, Department of Pharmacology, Yale University (1989.06-1991.09)
1.Functional study of MALT1.
2.Study on anti-HBV agents.
Research OutlineMucosa-associated-lymphoid-tissue (MALT) lymphoma of stomach is the most common extranodal lymphomas of human. Gastric MALT lymphoma is generally associated with H. pylori; and the eradication of this gastric infection results in durable tumor remission in 70% of the patients. Early identification of those patients in whom this approach is unlikely to be successful is a challenge to the clinicians. We, along with several distinguished groups, found that aberrant nuclear BCL10 expression is predictive of H. pylori-independent status in gastric MALT lymphoma. BCL10 normally resides in the cytoplasm and participates in signals to activate NF-B. The activation of NF-B is often associated with BCL10 phosphorylation. We started with the intentions listed below.
1. elucidating mechanism(s) involved in mediating the aberrant nuclear BCL10 expression.
Evidences indicated that aberrant nuclear localization of BCL10 might simply be due to interruption of its ability to self-associate or interact with cytoplasmic partners such as MALT1. In the process of studying the interaction of BCL10 and MALT1, BCL10 was found to be processed to a truncated form by MALT1. The findings lead us to the proposed studies thereafter.
2. identification of the phosphorylation site(s) of BCL10 and unraveling the myth of BCL10 phosphorylation.
Overexpression of BCL10 induced BCL10 phosphorylation, NF-B activation and the appearance of filamentous structure under microscope. BCL10L41R, a CARD mutated BCL10, lost all these BCL10-induced abilities. Neither N’BCL10GFP (1~114 a.a.) nor C’BCL10GFP (114~233 a.a.) was phosphorylated, indicating that both CARD and Serine/Threonine rich domain are required for BCL10 phosphorylation. Though not phosphorylated, N’BCL10GFP retained the ability to activate NF-B, suggesting that BCL10 phosphorylation was not essential for NF-B activation. The level of BCL10 phosphorylation was diminished by dominant-negative IκB through blocking NF-B activation.
Site directed mutagenesis was utilized to generate BCL10 mutants to map the phosphorylation sites on BCL10. In addition to the reported Serine 134,136,138,141 and 144, Serine 170 and 171 were also found to be major phosphorylation sites. Mutation with serine to alanine at these sites completely abolished the appearance of highly-molecular-weight phosphorylation isoforms. However, the un-phosphorylated BCL10 mutant retained its ability to activate NF-B and formed filamentous structure upon overexpression in 293T cells as wild type BCL10.
3. studying the nature of BCL10 cleavage
Arginine 228 was the reported cleavage site. As expected, BCL10R228G could not be cleaved by MALT1. However, in our assay system, deletion of Leucine225 abolished the nature of BCL10 being a substrate of MALT1. Site-directed mutagenesis was employed to investigate the role of amino acid residue Leu225 of BCL10 played as a substrate of MALT1. MALT1 failed to process BCL10L225A, BCL10L225G, BCL10L225Q, BCL10L225E mutants. But Mutants BCL10L225K and BCL10L225R could be processed by MALT1 as wild type BCL10. Interestingly, double mutants BCL10L225R,R228G retained its ability to be cleaved by MALT1. (The study has resulted in the publication of the following paper : J Biomed Sci. 2012; 19:85).
4. functional consequences of MALT1-mediated BCL10 cleavage
Truncated BCL10 induced NF-B activation and the appearance of filamentous structure under microscope, in analogy to wild type BCL10. Though phosphorylated, truncated BCL10 did display a different phosphorylation pattern as visualized by the different mobility of the phosphorylated products.
5. searching for physiological conditions that would trigger the cleavage of BCL10
Multiple cell lines including：Jurkat, Raji, Akata, NK, Raw264.7, THP-1, HeLa, AGS, and 293T were treated with stimulators, such as PMA/ionomycin, TNF-, LPS, Zymosan, IL-1, Helicobacter pylori, known to activate NF-B signaling. The levels of IB were examined as the indicator for NF-B activation. Cleavage of BCL10 or not could easily be monitored by denaturing polyacrylamide gel electrophoresis. Cleavage of BCL10 was observed only in cells（e.g. Jurkat, Raji, Akata, NK）of lymphoid origin in response to PMA/ionomycin treatment. Proteasome inhibitor, MG132, had no effect on the cleavage of BCL10. Bay 11-7085, IKK inhibitor, attenuated the level of BCL10 cleavage. Cytosolic calcium chelator, BAPTA, also downregulated the PMA/ionomycin-induced BCL10 cleavage. Taking together, IKK activity and Calcium might play certain roles in the cleavage of BCL10 in lymphoid cells.
6. generating constitutively-active paracaspase by fusion caspase-like domain of MALT1 with oligomierization motif from different molecules
Different fusion constructs （C106-2IgCLD，G-2IgCLD，and IAP2-2IgCLD）with oligomerization domain from caspase recruitment domain (CARD) of BCL10, dimerization domain of E. coli gyrase B and BIR domain of IAP2 were tested and compared for their abilities to activate NF-B or proteolytically process BCL10 substrate. Fusion construct C106-2IgCLD showed proteolytic activity but with minimum NF-B activation activity. G-2IgCLD showed potent NF-B activation activity but no proteolytic activity. IAP2-2IgCLD showed both potent NF-B activation and proteolytic activity. Fusion constructs with different oligomerization moiety showed apparently distinct properties. (Manuscript in preparation)
Publications 30 2015-12-10 09:33:50
Ordered by publish date
- Maintenance of Epstein-Barr virus latent status by a novel mechanism, latent membrane protein 1-induced interleukin-32, via the protein kinase Cδ pathway Journal of Virology 2015 | journal-article vol.89,no.11,page.5968-5980 Scopus: 8 Web of Science: 8 Impact Factor: 4.663
- Glycogen synthase kinase 3 negatively regulates IFN regulatory factor 3 transactivation through phosphorylation at its linker region Innate Immunity 2014 | journal-article vol.20,no.1,page.78-87 Scopus: 7 Web of Science: 5 Impact Factor: 2.342
- BCL10GFP fusion protein as a substrate for analysis of determinants required for Mucosa-Associated Lymphoid Tissue 1 (MALT1)-mediated cleavage Journal of Biomedical Science 2012 | journal-article vol.19,no.1 Scopus: 0 Web of Science: 0 Impact Factor: 2.799
- Epstein-barr virus BGLF4 kinase downregulates NF-κB transactivation through phosphorylation of coactivator UXT Journal of Virology 2012 | journal-article vol.86,no.22,page.12176-12186 Scopus: 14 Web of Science: 14 Impact Factor: 4.663
- Involvement of recepteur d'origine nantais receptor tyrosine kinase in Epstein-Barr virus-associated nasopharyngeal carcinoma and its metastasis American Journal of Pathology 2012 | journal-article vol.181,no.5,page.1773-1781 Scopus: 8 Web of Science: 6 Impact Factor: 4.057
- Characterization of Epstein - Barr virus BGLF4 kinase expression control at the transcriptional and translational levels Journal of General Virology 2010 | journal-article vol.91,no.9,page.2186-2196 Scopus: 4 Web of Science: 4 Impact Factor: 2.838
- Epstein-barr virus BGLF4 kinase suppresses the interferon regulatory factor 3 signaling pathway Journal of Virology 2009 | journal-article vol.83,no.4,page.1856-1869 Scopus: 63 Web of Science: 61 Impact Factor: 4.663
- Differential response to H. pylori eradication therapy of co-existing diffuse large B-cell lymphoma and MALT lymphoma of stomach - Significance of tumour cell clonality and BCL10 expression Journal of Pathology 2007 | journal-article vol.211,no.3,page.296-304 Scopus: 13 Web of Science: 11 Impact Factor: 6.894
- Differential expression of osteoblast-specific factor 2 and polymeric immunoglobulin receptor genes in nasopharyngeal carcinoma Head and Neck 2005 | journal-article vol.27,no.10,page.873-882 Scopus: 23 Web of Science: 22 Impact Factor: 3.376
- Expression of CD86 and increased inflitration of NK cells are associated with Helicobacter pylori-dependent state of early stage high-grade gastric MALT lymphoma World Journal of Gastroenterology 2005 | journal-article vol.11,no.28,page.4357-4362
- Nuclear expression of BCL10 or nuclear factor kappa B helps predict Helicobacter pylori-independent status of low-grade gastric mucosa-associated lymphoid tissue lymphomas with or without t(11;18)(q21;q21 ) Blood 2005 | journal-article vol.106,no.3,page.1037-1041 Scopus: 57 Web of Science: 50 Impact Factor: 13.164
- Inhibition of the Epstein-Barr virus lytic cycle by Zta-targeted RNA interference Journal of General Virology 2004 | journal-article vol.85,no.6,page.1371-1379 Scopus: 41 Web of Science: 40 Impact Factor: 2.838
- Nuclear expression of BCL10 or nuclear factor kappa B predicts Helicobacter pylori-independent status of early-stage, high-grade gastric mucosa-associated lymphoid tissue lymphomas Journal of Clinical Oncology 2004 | journal-article vol.22,no.17,page.3491-3497 Scopus: 55 Web of Science: 48 Impact Factor: 24.008
- Hepatitis B virus X protein activates a survival signaling by linking Src to phosphatidylinositol 3-kinase Journal of Biological Chemistry 2003 | journal-article vol.278,no.34,page.31807-31813 Scopus: 48 Web of Science: 43 Impact Factor: 4.125
- Thyroid hormone receptor α 1 (c-erb A α 1) suppressed transforming phenotype of nasopharyngeal carcinoma cell line Cancer Letters 2002 | journal-article vol.184,no.2,page.149-156 Scopus: 7 Web of Science: 5 Impact Factor: 6.375
- Coumarins and antiplatelet constituents from the root bark of Zanthoxylum schinifolium Planta Medica 2000 | journal-article vol.66,no.7,page.618-623 Scopus: 52 Web of Science: 51 Impact Factor: 2.342
- Elevated expression of thyroid hormone receptor α2 (c-erb A-α2) in nasopharyngeal carcinoma British Journal of Cancer 2000 | journal-article vol.83,no.12,page.1674-1680 Scopus: 6 Web of Science: 6 Impact Factor: 6.176
- Hepatitis b virus x protein inhibits transforming growth factor-β-induced apoptosis through the activation of phosphatidylinositol 3-kinase pathway Journal of Biological Chemistry 2000 | journal-article vol.275,no.33,page.25858-25864 Scopus: 157 Web of Science: 147 Impact Factor: 4.125
- Oltipraz, a novel inhibitor of hepatitis B virus transcription through elevation of p53 protein Carcinogenesis 1998 | journal-article vol.19,no.12,page.2133-2138 Scopus: 18 Web of Science: 23 Impact Factor: 5.105
- Transactivation of the human MDR1 gene by hepatitis B virus X gene product Journal of Hepatology 1998 | journal-article vol.29,no.6,page.872-878 Scopus: 11 Web of Science: 10 Impact Factor: 12.486
- Coumarins and anti-HBV constituents from Zanthoxylum schinifolium Phytochemistry 1997 | journal-article vol.45,no.7,page.1419-1422 Scopus: 57 Web of Science: 47 Impact Factor: 3.205
- Tax of the human T-lymphotropic virus type I transactivates promoter of the MDR-1 gene Biochemical and Biophysical Research Communications 1997 | journal-article vol.238,no.2,page.482-486 Scopus: 8 Web of Science: 9 Impact Factor: 2.466
- Differential display and cloning of messenger RNA from human normal nasal epithelial cells versus nasopharyngeal carcinoma cell lines. Zhonghua Minguo wei sheng wu ji mian yi xue za zhi = Chinese journal of microbiology and immunology 1996 | journal-article vol.29,no.4,page.225-231
- Characterization of conditions for the activation of endogenous guinea pig retrovirus in cultured cells by 5-bromo-2′-deoxyuridine Virus Genes 1995 | journal-article vol.9,no.3,page.201-209 Scopus: 3 Web of Science: 3 Impact Factor: 1.431
- Effect of anti-HIV 2′-β-fluoro-2′,3′-dideoxynucleoside analogs on the cellular content of mitochondrial DNA and on lactate production Biochemical Pharmacology 1994 | journal-article vol.48,no.7,page.1477-1481 Scopus: 34 Web of Science: 35 Impact Factor: 4.581
- Deoxycytidine deaminase-resistant stereoisomer is the active form of (±)-2′,3′-dideoxy-3′-thiacytidine in the inhibition of hepatitis B virus replication Journal of Biological Chemistry 1992 | journal-article vol.267,no.20,page.13938-13942
- Erratum: Deoxycytidine deaminase-resistant stereoisomer is the active form of (±)-2',3'-dideoxy-3'-thiacytidine in the inhibition of hepatitis B virus replication (Journal of Biological Chemistry (1992) 267 (13938-13942)) Journal of Biological Chemistry 1992 | journal-article vol.267,no.33
- Synthesis of enantiomerically pure (2′R,5′S)-(-)-1-[2-(hydroxymethyl)oxathiolan-5-yl]cytosine as a potent antiviral agent against Hepatitis B Virus (HBV) and Human Immunodeficiency Virus (HIV) Journal of Organic Chemistry 1992 | journal-article vol.57,no.8,page.2217-2219
- Inhibition of the replication of hepatitis B virus in vitro by 2′,3′-dideoxy-3′-thiacytidine and related analogues Proceedings of the National Academy of Sciences of the United States of America 1991 | journal-article vol.88,no.19,page.8495-8499
- 5-Fluorouracil substitution alters pre-mRNA splicing in vitro Journal of Biological Chemistry 1988 | journal-article vol.263,no.9,page.4467-4473
Projects 21 2015-12-10 10:01:48
Ordered by project period
Studies on the Functional Consequences of Autocleavage of Human Paracaspase-MALT1 (人類Paracaspase-MALT1 自我切割生物功能的探討)Project Number : NSC102-2320-B002-024 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 10208~10307
Studies on the Biological Consequences and Mechanism(s) Involved in the Cleavage of BCL10 (引起BCL10蛋白質切割的機轉探討及生物意義研究)Project Number : NSC97-2320-B002-011-MY3 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 9908~10007
Studies on the Biological Consequences and Mechanism(s) Involved in the Cleavage of BCL10 (引起BCL10蛋白質切割的機轉探討及生物意義研究)Project Number : NSC97-2320-B002-011-MY3 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 9808~9907
Studies on the Biological Consequences and Mechanism(s) Involved in the Cleavage of BCL10 (引起BCL10蛋白質切割的機轉探討及生物意義研究)Project Number : NSC97-2320-B002-011-MY3 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 9708~9807
Examination on Microrna Signatures Associated with Mucosa-Associated Lymphoid Tissue (MALT) B-Cell Lymphomas (檢視和黏膜相關性淋巴組織淋巴癌(MALT Lymphoma) 有關的MicroRNA表現標記)Project Number : NSC95-2320-B002-090 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : 鄭安理 Funding IC : 行政院國家科學委員會 Project Period : 9508~9607
Effects of BCL10 Phosphorylation on Its Function and Cellular Localization (BCL10蛋白質磷酸化對其功能及細胞位移影響的研究)Project Number : NSC94-2320-B002-069 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : 鄭安理 Funding IC : 行政院國家科學委員會 Project Period : 9408~9507
Studies on the Aberrant Nuclear BCL10 Expression in Mucosa-Associated Lymphoid Tissue (MALT) (在黏膜相關性淋巴組織淋巴癌中不正常細胞核內BCL10表現的研究)Project Number : NSC93-2320-B002-075 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : 鄭安理 Funding IC : 行政院國家科學委員會 Project Period : 9308~9407
Study on Modulation of Hepatitis B Virus Replication and/or Infectin by Small RNAs (利用小RNAs控制B型肝炎病毒複製和感染的研究)Project Number : NSC92-2320-B002-186 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 9208~9307
Utilization of SiRNAs for Regulation of Gene Expression in Hepatoma Cell Lines (利用siRNAs在肝癌細胞株中調控基因表現)Project Number : NSC91-2320-B002-190 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 9108~9207
Roles of Specific Cellular Factors with Differential Expressino Pattern in the Carcinogenesis Process of Nasopharyngeal Carcinoma (EB病毒與細胞因子在鼻咽癌癌化過程角色之探討(III)---探討鼻咽癌細胞中特異性細胞因子在癌化過程扮演的角色)Project Number : NSC90-2320-B002-182 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 9008~9107
Elucidation of Mechanism(s) Involved in the Attenuation of Colony Formation in Hepatoma Cells by Hepatitis B Virus X Protein (B型肝炎病毒X蛋白質降低細胞群落形成機轉的探討)Project Number : NSC90-2320-B002-142 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 9008~9107
Roles of Specific Cellular Factors with Differential Expressino Pattern in the Carcinogenesis Process of Nasopharyngeal Carcinoma (EB病毒與細胞因子在鼻咽癌癌化過程角色之探討(II)---探討鼻咽癌細胞中特異性細胞因子在癌化過程扮演的角色)Project Number : NSC89-2320-B002-207 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8908~9007
Elucidation of the Mechanism(s) Involved in the Activation of Phosphatidyllinositol-3-kinase by Hepatitis B Virus X Protein (B型肝炎病毒X蛋白質活化Phosphatidylinositol-3-Kinase機轉的探討)Project Number : NSC89-2320-B002-194 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8908~9007
Elucidation of the Mechanism(s) Involved in the Activation of Phosphatidyinositol-3-Kinase by Hepatitis B Virus X Protein (B型肝炎病毒X蛋白質活化Phosphatidyinositol-3-Kinase機轉的探討)Project Number : NSC89-2320-B002-033 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8808~8907
Roles of Specific Cellular Factors with Differential Expressino Pattern in the Carcinogenesis Process of Nasopharyngeal Carcinoma (EB病毒與細胞因子在鼻咽癌癌化過程角色之探討(I)---探討鼻咽癌細胞中特異性細胞因子在癌化過程扮演的角色)Project Number : NSC89-2320-B002-048 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8808~8907
Cloning and Functional Analysis of a Gene Expressed in Normal Nasal Epithelial Cells and Not in Nasopharyngeal Carcinoma Cell Line (選殖及分析一個只在正常鼻腔上皮細胞而不在鼻咽癌細胞株表現的基因)Project Number : NSC88-2314-B002-153 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8708~8807
Cloning and Functional Analysis of a Gene Expressed in Normal Nasal Epithelial Cells and Not in Nasopharyngeal Carcinoma Cell Line (鼻咽癌與EB病毒之研究---擇殖及分析一個只在正常鼻腔上皮細胞，而不在鼻咽癌細胞株表現的基因)Project Number : NSC87-2314-B002-268 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8608~8707
Cloning and Functional Analysis of a Gene Expressed in Normal Nasal Epithelial Cells and Not in Nasopharyngeal Carcinoma Cell Line (鼻咽癌與EB病毒之研究---擇殖及分析一個只在正常鼻腔上皮細胞，而不在鼻咽癌細胞株表現的基因)Project Number : NSC86-2314-B002-266 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8508~8607
Studies of Hepatitis B Virus X Gene Product on Inhibition of Cellular Apoptosis and Induction of Genomic Instability Phenotype (肝炎研究---B型肝炎病毒X蛋白防細胞自戕及導致基因不穩定可能性的探討)Project Number : NSC86-2315-B002-013 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8508~8607
Studies on the Identification of Cellular Factor(s) that Mediate-HBV X Gene Productaction (探索引導B型肝炎病毒X蛋白質動作之細胞因子的研究)Project Number : NSC85-2331-B002-074 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : Funding IC : 行政院國家科學委員會 Project Period : 8408~8507
Biological Studies of Guinea Pig Endogenous Retrovirus (天竺鼠內生性反轉錄病毒生物活性的探討)Project Number : NSC82-0412-B002-533 PI : SHIN-LIAN DOONG (董馨蓮) Co-PI : 林逢祺 Funding IC : 行政院國家科學委員會 Project Period : 8202~8301