Developmental Dyslexia (DD) refers to a reading disorder affecting individuals that possess otherwise normal intelligence. Having demonstrated by familial and twin studies, genetic factors are found to be of major significance to DD development. A strong dyslexia susceptibility gene KIAA0319 (K), of which crucial role in DD had been revealed by various linkage and association studies, was found to have 40% reduction in expression in the DD risk haplotype. Besides, both up- and down-regulation of K would result in impaired neuronal migration in rat. Despite the undoubtedly strong linkage of K to DD, biological and molecular knowledge of K is still lacking. Consequently, how K plays its role in DD remains unclear. To address this question, investigations of human K protein and its interactions in molecular level were performed. K protein is a large transmembrane protein which consists of four main parts, including the N-terminus of K which has a MANSC domain downstream of the signal peptide, a large cluster of five PKD domains in the middle of the protein sequence, a Cysteine -rich C6 region together with a transmembrane domain which had been demonstrated to be critical for forming K protein homodimer, and the only cytoplasmic C-terminus of K. Having shown that no gross effect on gene expression at both mRNA and protein level was found with overexpressing K by DNA microarray and two-dimensional gel electrophoresis, protein interactions involving K were targeted for investigation. Towards this goal, a monoclonal antibody against K was raised, which is capable for recognizing native full-length K proteins in immunoblotting, indirect immunofluorescence staining, as well as in immunoprecipitation. A novel K interaction partner protein KIAA0319-Like (KL), which is a homologous protein of K with high sequence similarity (59%), has been found and confirmed by co-immunoprecipitation. No interaction was shown for truncation mutants of Cysteine-rich C6 region in either K or KL proteins, cuing that the interaction of K and KL at C6 region is a mimic of K homodimer, and led to a hypothesis that the function of K is regulated by KL, which serves as a molecular control of neuronal migration by regulating the formation of K dimer. Another known interaction partner of K protein, the mu---subunit of Adaptor protein 2 complex (AP2M1) which binds to cytoplasmic C-terminus of K (55% similarity to that of KL), was found to have similar binding behaviour towards K as well as KL by co-immunoprecipitation and molecular docking. In addition to AP2M1, two adaptor proteins FEM and SH2 were also confirmed to be interacting with cytoplasmic C-terminus of K, suggested that cytoplasmic region of K is responsible for interactions of downstream cellular pathways. Interaction of K with adaptor proteins also suggested that K might be a membrane receptor that mediates signalling via various adapter proteins. The N-terminus of K protein which has the least sequence similarity to KL (31%) is hence thought to confer to the specificity of the receptor and is critical to the function of K in DD. / Chan, Hoi Ling. / Adviser: Mary M. Y. Waye. / Source: Dissertation Abstracts International, Volume: 72-04, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 164-170). / 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.

Wong, Chi Bun. / Thesis submitted in: Nov 2006. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 116-127). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.ii / 摘要 --- p.v / Abbreviations --- p.vii / List of Table and Figures --- p.ix / Table of Contents --- p.x / Chapter Chapter 1 --- p.1 / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Identification of the proteins regulated by BRE when BRE was over-expressed or silencedin C2C12 and D122 --- p.1 / Chapter 1.1.1 --- What is BRE? --- p.1 / Chapter 1.1.2 --- BRE gene is Highly Conserved --- p.2 / Chapter 1.1.3 --- BRE binds to the Intracellular Domain of TNFR1 and Fas --- p.3 / Chapter 1.1.4 --- BRE Suppresses Apoptosis --- p.4 / Chapter 1.1.5 --- "BRE forms a Holoenzyme Complex with BRCA1, BARD1 and BRCC36" --- p.4 / Chapter 1.16 --- Roles of the Differentially Expressed Proteins Identified in the siRNA knockdown Experiments --- p.5 / Chapter 1.1.6.1 --- Akt3 --- p.5 / Chapter 1.1.6.2 --- Mdm2/4 --- p.6 / Chapter 1.1.6.3 --- Prohibitin --- p.7 / Chapter 1.1.6.4 --- Carbonic Anhydrase III --- p.8 / Chapter 1.1.6.5 --- 26S Proteasome --- p.8 / Chapter 1.2 --- The Role of BRE in Liver: a morphological approach --- p.9 / Chapter 1.2.1 --- The General Structure of the Liver. --- p.9 / Chapter 1.2.2 --- The Essential Functions of the Liver --- p.11 / Chapter 1.2.3 --- Inflammation of the Liver --- p.11 / Chapter 1.2.3.1 --- Hepatitis --- p.11 / Chapter 1.2.3.2 --- Acute Hepatitis --- p.12 / Chapter 1.2.3.3 --- Chronic Hepatitis --- p.12 / Chapter 1.2.4 --- Necrosis and Apoptosis --- p.13 / Chapter 1.2.5 --- The Apoptotic Pathway --- p.14 / Chapter 1.2.6 --- Hepatic Necrosis is Divided into Different Zones --- p.16 / Chapter 1.2.6.1 --- Hepatitis Necrosis is Categorized into 3 Zones --- p.16 / Chapter 1.2.7 --- Carbon Tetrachloride (CCL4) --- p.16 / Chapter 1.2.8 --- TNFa is a Pleiotropic Cytokine --- p.17 / Chapter 1.3 --- The Objectives of This Project --- p.20 / Chapter Chapter 2 --- p.21 / Chapter 2. --- Materials and Methods --- p.21 / Chapter 2.1 --- Animals --- p.21 / Chapter 2.2 --- Adminstration of Carbon Tetrachloride and Corn Oil --- p.21 / Chapter 2.3 --- Cell Cultures --- p.22 / Chapter 2.4 --- Cell Culturing --- p.22 / Chapter 2.5 --- Gene Silencing with Small Interfering RNA (siRNA) --- p.23 / Chapter 2.5.1 --- Transfection with BRE siRNA --- p.24 / Chapter 2.6 --- Cell Proliferation Assays --- p.24 / Chapter 2.7 --- In-Situ Hybridization of BRE Sense and Antisense Probes --- p.25 / Chapter 2.8 --- Immunohistological Staining --- p.26 / Chapter 2.9 --- Semi-Quantitative RT-PCR --- p.28 / Chapter 2.10 --- Comparative Proteomics --- p.29 / Chapter 2.10.1 --- Sample Preparation for Two Dimensional Gel Electrophoresis --- p.29 / Chapter 2.10.2 --- Two Dimensional Polyacrylamide Gel Electrophoresis --- p.30 / Chapter 2.10.3 --- In-Gel Digestion and MALDI-TOF Analysis --- p.31 / Chapter 2.11 --- Western Blotting --- p.32 / Chapter 2.12 --- Flow Cytometry --- p.34 / Chapter 2.13 --- Haematoxylin and Eosin Staining (H&E) --- p.34 / Chapter Chapter 3 --- p.36 / Chapter 3. --- Results --- p.36 / Chapter 3.1 --- BRE expression in C2C12 cells --- p.36 / Chapter 3.2 --- Comparative Proteomic Profile of BRE silenced C2C12 cells --- p.41 / Chapter 3.3 --- Effect of Silencing BRE on C2C12 cell Proliferation --- p.49 / Chapter 3.4 --- Effects of BRE over-expression in D122 cells --- p.54 / Chapter 3.5 --- BRE Expression in the Liver --- p.62 / Chapter 3.5.1 --- Histological Analysis of Liver Sections after 24 hours of CCL4 Insult --- p.62 / Chapter 3.5.2 --- BRE Expression in the Liver --- p.62 / Chapter 3.6 --- Histological Study of Liver Treated with CCL4 --- p.67 / Chapter 3.7 --- BRE Expression in Experimental Liver --- p.76 / Chapter Chapter 4 --- p.92 / Chapter 4. --- Discussion --- p.92 / Chapter 4.1 --- Expression of BRE in C2C12 --- p.92 / Chapter 4.2 --- The Regulatory Function of BRE --- p.96 / Chapter 4.3 --- The Relationship Between BRE and p53 --- p.98 / Chapter 4.4 --- The Relationship Between BRE and NFkB --- p.104 / Chapter 4.5 --- BRE Expression in Normal Control and CCL4 Treated Livers --- p.105 / Chapter 4.6 --- A Possible Explanation for the Necrosis Pattern Observed --- p.107 / Chapter 4.7 --- The Relationship Between BRE and the TNF Receptors --- p.109 / Chapter Chapter 5 --- p.112 / Chapter 5. --- Conclusion and Future Prospects --- p.112 / References --- p.116

Wong, Wan Man. / Thesis submitted in: December 2006. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 85-98). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract in Chinese --- p.iii / Acknowledgements --- p.v / Lists of Figures and Tables --- p.vi / Table of Abbreviations --- p.xi / Table of Contents --- p.xv / Chapter Chapter I --- Introduction / Chapter 1.1 --- Brain and Reproductive Organ Expressed Gene --- p.1 / Chapter 1.2 --- Programmed cell death --- p.4 / Chapter 1.3 --- Limb development in mouse --- p.8 / Chapter 1.4 --- Role of BRE in apoptosis --- p.12 / Chapter 1.5 --- Role of programmed cell death in interdigital tissue regression --- p.14 / Chapter 1.6 --- Aim of study --- p.17 / Chapter Chpater II --- Materials and methods / Chapter 2.1 --- Mice --- p.18 / Chapter 2.2 --- In-situ hybridization / Chapter 2.2.1 --- Histology --- p.18 / Chapter 2.2.2 --- Preparation of riboprobe for in-situ hybridization --- p.19 / Chapter 2.2.3 --- In-situ hybridization --- p.20 / Chapter 2.3 --- Interdigital tissue culture --- p.21 / Chapter 2.4 --- Gene interference / Chapter 2.4.1 --- Construction of Bre-siRNA --- p.22 / Chapter 2.4.2 --- siRNA transfection of cultured interdigital cells --- p.23 / Chapter 2.5 --- Semi-quantitative RT-PCR / Chapter 2.5.1 --- Sample collection of interdigital cells and explants --- p.23 / Chapter 2.5.2 --- RNA isolation and extraction --- p.24 / Chapter 2.5.3 --- Reverse-transcription and cDNA synthesis --- p.25 / Chapter 2.5.4 --- Polymerase chain reaction --- p.26 / Chapter 2.6 --- Assay of cell viability by MTT --- p.28 / Chapter 2.7 --- Comparative proteomics --- p.30 / Chapter 2.7.1 --- Collection of interdigital cells --- p.30 / Chapter 2.7.2 --- Preparation of cell lysate --- p.31 / Chapter 2.7.3 --- Assay of protein concentration in cell lysate --- p.31 / Chapter 2.7.4 --- Two-dimensional gel electrophoresis --- p.33 / Chapter 2.7.5 --- Protein identification by mass fingerprinting --- p.36 / Chapter 2.8 --- Statistical Method --- p.38 / Chapter Chapter III --- Results / Chapter 3.1 --- Spatial and temporal expression of Bre in murine embryonic hindlimbs --- p.39 / Chapter 3.2 --- Expression of Bre isoforms in interdigital tissues --- p.45 / Chapter 3.3 --- Silencing of Bre expression by siRNA in interdigital cells --- p.49 / Chapter 3.4 --- Effect on viability of Bre-silenced interdigital cells by siRNA --- p.51 / Chapter 3.5 --- Comparative proteomic profile of Bre-silenced interdigital cultured cells --- p.53 / Chapter 3.6 --- Identification of proteins that were differentially expressed by MALDI- TOF --- p.71 / Chapter 3.7 --- The mRNA levels of proteins identified that were differentially expressed --- p.74 / Chapter Chapter IV --- Discussion --- p.77 / References --- p.85 / Appendices --- p.99 / Publication --- p.108

Li Qing. / "July 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 161-192). / 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.