Brefeldin A (BFA) has been a useful tool for studying organelle dynamics and protein trafficking in plant cells. Using several Golgi (MAN1 and GONST1) and TGN (SCAMP1 and SYP61) fluorescent protein markers as tools, I have showed that BFA-induced aggregates from Golgi apparatus and TGN are morphologically distinct in the same plant cells. In addition, the internalized endosomal marker FM4-64 colocalized with the TGN-derived aggregates but separated from the Golgi aggregates. In the presence of the endocytosis inhibitor tyrphostin A23, SCAMP1 and FM4-64 are largely excluded from the TGN SYP61-positive BFA-induced aggregates, indicating homotypic fusion of TGN rather than de novo endocytic trafficking is important for the formation of TGN/EE-derived BFA-induced aggregates. Since the TGN also serves as an EE receiving materials from plasma membrane continuously, these data therefore support the notion that the secretory Golgi organelle is distinct from the endocytic TGN/EE in response to BFA treatment in plant cells. / Little is known about the trafficking mechanism of plasma membrane (PM) proteins in the endomembrane system of plant cells that contain several membrane-bound organelles including the endoplasmic reticulum (ER), Golgi, trans-Golgi network (TGN) of early endosome (EE), prevacuolar compartment (PVC) or late endosome (LE). Here, I study the transport pathway and sorting signals of secretory carrier membrane protein 1 (SCAMP1) by following its transient expression in tobacco BY-2 protoplasts and show that SCAMP1 reaches the PM via an ER-Golgi-TGN-PM pathway. Loss-of-function and gain-of-function analysis of various GFP fusions with SCAMP1 mutations further demonstrates that: (1) the cytosolic N terminus of SCAMP1 contains an ER export signal; (2) the transmembrane domain 2 (TMD2) and TMD3 of SCAMP1 are essential for Golgi export; and (3) SCAMP1 TMD1 is essential for TGN-to-PM targeting. Therefore, both the cytosolic N-terminus and TMD sequences of SCAMP1 play integral roles in mediating its transport to the PM via an ER-Golgi-TGN pathway. / Cai, Yi. / Adviser: Liwen Jiang. / Source: Dissertation Abstracts International, Volume: 73-02, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 93-102). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Shan Sze Wan. / "July 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 125-139) / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Cancer/testis (CT) antigens are encoded by genes that are normally expressed only in the human germ-line, but are also expressed in various tumor types. CT antigens are also being studied for their roles in carcinogenesis as well as for their potentials as targets for anti-cancer therapy. A novel CT gene, NYD-SP8, (Accession No. AY014285.1) has recently been identified. It is located to human chromosome 19q13.31 and encodes a 27 kDa glucosylphosphatidylinositol (GPI) anchored cell surface protein, which shows structural homology to urokinase plasminogen activator receptor (uPAR). This thesis describes the characterization and functional roles of NYD-SP8 involved in cancer development. / In summary, the present findings have demonstrated the roles of NYD-SP8 in multi-step cancer development. Further investigations of NYD-SP8 in cancer development may provide new insights and ground for potential use of CT antigens in anti-cancer therapy. (Abstract: 428 words) / In the first set of experiments, the possible role(s) and underlying mechanism(s) of NYD-SP8 in regulating cell proliferation and apoptosis were investigated. Flow cytometric analysis, cell proliferation assay and Western blot analysis showed that NYD-SP8 promoted cell proliferation and protected cells against TNFalpha-induced apoptosis in Human embryonic kidney cells (HEK293) and human hepatocellular carcinoma cells (hHCC). In vitro studies showed that NYD-SP8 enhanced anchorage-independent growth of hHCC, further suggesting the pro-survival effect of NYD-SP8. These data demonstrated important functions of NYD-SP8 in promoting cell growth and preventing apoptosis during cancer development. / In the last part of thesis, the involvement of NYD-SP8 in epithelial-mesenchymal transitions (EMTs) was demonstrated. Upon TGFbeta stimulation or TGFbeta/TNFalpha co-stimulation, the mRNA and protein expression of NYD-SP8 was decreased in LIM1863 cells. Cell adhesion assay showed that the attachment ability of hHCC-SP8 was lowered in laminin and fibronectin coated plate, suggesting the possible role of NYD-SP8 in affecting cell-matrix interaction. These data indicate that NYD-SP8 is involved in the EMTs process and may serve as potential EMTs markers during cancer development. / In the second sets of experiments, the possible role(s) and underlying mechanism(s) of NYD-SP8 in regulating cancer invasion and metastasis were investigated. The results showed that NYD-SP8 could suppress multiple "tumor associated" proteases. Overexpression of NYD-SP8 resulted in reducing activities of the three major classes of proteases known to be involved in ECM degradation, including uPA, matrix metalloproteinases (MMPs) and cathepsin B, leading to suppression of both in vitro and in vivo cancer cell invasion and metastasis. Co-immunoprecipitation experiments showed binding of NYD-SP8 to uPA/uPAR complexes and interfering with active uPA production. These data demonstrated an important function of NYD-5P8 in regulating ECM degradation, providing a novel mechanism that modulates uPA/uPAR signaling in the suppression of cancer progression. / Chung, Chin Man. / "December 2009." / Adviser: H.C. Chan. / Source: Dissertation Abstracts International, Volume: 72-01, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 124-150). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

San, Wan Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 75-78). / Abstracts in English and Chinese. / Thesis/Assessment Committee --- p.i / Statement --- p.ii / Abstract --- p.iii / 摘要 --- p.v / Acknowledgements --- p.vi / Table of Contents --- p.viii / List of Tables --- p.x / List of Figures --- p.xi / List of Abbreviations --- p.xii / Chapter Chapter 1 --- General Introduction --- p.1 / Chapter 1.1 --- The Plant Secretory Pathway --- p.1 / Chapter 1.2 --- AtEMP70 As a Potential Candidate in PVC Proteomics Analysis --- p.4 / Chapter 1.3 --- EMP70 Protein Family --- p.6 / Chapter 1.3.1 --- Arabidopsis EMP70 Protein Family --- p.6 / Chapter 1.3.2 --- EMP70 Homologs Among Different Species --- p.9 / Chapter 1.4 --- Aims of This Study --- p.10 / Chapter Chapter 2 --- Materials and Methods --- p.12 / Chapter 2.1 --- Generation of Arabidopsis cDNA --- p.12 / Chapter 2.2 --- Plasmid Construction --- p.13 / Chapter 2.3 --- Transformation of Tobacco BY-2 Cells --- p.14 / Chapter 2.4 --- Confocal Immunofluorescence Studies --- p.15 / Chapter 2.5 --- Drug Treatments --- p.16 / Chapter 2.6 --- Transient Expression in Protoplasts --- p.16 / Chapter 2.7 --- Generation of Antibodies --- p.18 / Chapter 2.8 --- SDS-PAGE and Western Blot Analysis --- p.19 / Chapter 2.9 --- Microsomal Protein Extraction --- p.21 / Chapter 2.10 --- Subcellular Fractionation --- p.21 / Chapter 2.11 --- Membrane Strip-off --- p.23 / Chapter Chapter 3 --- Results --- p.24 / Chapter 3.1 --- Subcellular Localization Study of GFP-tagged AtEMP2 Fusions via Transient Expression --- p.24 / Chapter 3.1.1 --- AtEMP2-GFP Localized to TGN in BY-2 Protoplasts --- p.24 / Chapter 3.1.2 --- AtEMP2-GFP Localized to TGN in Arabidopsis Protoplasts --- p.30 / Chapter 3.1.3 --- N-terminal GFP-tagged AtEMP2 Fusions Localized to the Golgi Apparatus in Arabidopsis Protoplasts --- p.33 / Chapter 3.2 --- Generation and Characterization of Transgenic Tobacco BY-2 Cells and Arabidopsis PSB-L Cells Expressing AtEMP2-GFP Fusion --- p.36 / Chapter 3.2.1 --- Subcellular Localization of AtEMP2-GFP Fusion in Transgenic BY-2 Cell Lines --- p.36 / Chapter 3.2.2 --- Subcellular Localization of AtEMP2-GFP Fusion in Transgenic Arabidopsis PSB-D Cell Lines --- p.39 / Chapter 3.3 --- Immunofluorescent Labeling Study --- p.41 / Chapter 3.3.1 --- ManI Antibodies Did Not Label the Punctate Organelles --- p.41 / Chapter 3.3.2 --- AtEMP2 Antibodies Labeled the Golgi Apparatus --- p.43 / Chapter 3.4 --- Generation of AtEMP70 Antibodies --- p.46 / Chapter 3.5 --- Western Blot Analysis --- p.50 / Chapter 3.5.1 --- Heat Treatment Caused Aggregation of AtEMP2-GFP Fusion Proteins --- p.51 / Chapter 3.5.2 --- Size Change of AtEMP2-GFP Fusion Proteins in Response to Heat Treatment --- p.52 / Chapter 3.5.3 --- Aggregation Formation of AtEMP2-T7 Fusion Proteins in 95°C --- p.56 / Chapter 3.5.4 --- Distribution of Endogenous AtEMP70 in Arabidopsis Wild Type Cells --- p.58 / Chapter 3.6 --- Subcellular Fractionation --- p.61 / Chapter 3.6.1 --- C-terminal GFP- or T7-tagged Fusion Affected the Subcellular Localization of AtEMP2 --- p.61 / Chapter 3.6.2 --- Endogenous AtEMP70 Localized to the Golgi Apparatus --- p.64 / Chapter Chapter 4 --- Discussion and Future Perspectives --- p.67 / Chapter 4.1 --- Discussion --- p.67 / Chapter 4.1.1 --- ER Export Signal in the Cytosolic Tail of AtEMP70 --- p.71 / Chapter 4.1.2 --- Potential Golgi Retention Signal in the Cytosolic Tail of AtEMP70 --- p.73 / Chapter 4.2 --- Future Perspectives --- p.74 / References --- p.75

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 89-111).

There is no abstract available for this thesis. / Department of Chemistry

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 3 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 5 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 4 uppsatser.