Rab23 plays a role in the pathophysiology of mesangial cells - A proteomic analysis

• Published in: Proteomics (Weinheim) 2011-02, Vol.11 (3), p.380-394
• Main Authors: Huang, Tzu-Hao, Ka, Shuk-Man, Hsu, Yu-Juei, Shui, Hao-Ai, Tang, Bor-Luen, Hu, Kuang-Yu, Chang, Junn-Liang, Chen, Ann
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Blotting, Western; Cell biology; Cell Proliferation; Cells, Cultured; Constitutively active; Dominant negative; Electrophoresis, Gel, Two-Dimensional; Fundamental and applied biological sciences. Psychology; Mesangial cells; Mesangial Cells - metabolism; Mice; Miscellaneous; Promoter Regions, Genetic - genetics; Proteins; Proteome - analysis; Proteome - metabolism; Proteomics; rab GTP-Binding Proteins - antagonists & inhibitors; rab GTP-Binding Proteins - genetics; rab GTP-Binding Proteins - metabolism; Rab23; RNA, Small Interfering - genetics; shRNA knockdown; Signal transduction; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• ISSN: 1615-9853; 1615-9861; 1615-9861
• DOI: 10.1002/pmic.201000165

Rab23, a novel member of the Rab family of small GTPases, has recently been identified in mesangial cells (MCs). Although Rab23 levels in MCs are associated with glomerular nephropathies, the exact physiological and pathological roles of Rab23 in MCs are unknown. In the present study, its roles in MCs were explored by performing proteomics and systems biology analyses in MCs after knockdown or overexpression of Rab23. Knockdown of Rab23 was achieved by transfecting MCs with a plasmid expressing short hairpin RNA against Rab23, while overexpression of Rab23 was accomplished by transfection with the wild‐type, dominant negative, and constitutively active Rab23 gene constructs. The effects of different levels of Rab23 activity on proteome of various biological pathways were investigated. Gel‐based proteomic approaches and systems biology tools, respectively, were used to identify the Rab23‐regulated proteins and the functional pathways. Proteomic analysis revealed the potential roles for Rab23 in multiple processes, including G‐protein signal transduction, transcription modulation, RNA stabilization, protein synthesis and degradation, cytoskeleton reorganization, anti‐oxidation and detoxification, circadian rhythm regulation and phagocytosis. Bioinformatics analyses showed that Rab23 impacts on multiple biological networks in MCs. These data may shed light on the roles of Rab23 in mesangiopathy or MC damage.