title: Translocation of a human focal adhesion LIM-only protein, FHL2, during myofibrillogenesis and identification of LIM2 as the principal determinants of FHL2 focal adhesion localization.
abstract: LIM domain proteins are found to be important regulators in cell growth, cell fate determination, cell differentiation, and remodeling of the cell cytoskeleton. Human Four-and-a-half LIM-only protein 2 (FHL2) is expressed predominantly in human heart and is only slightly expressed in skeletal muscle. Since FHL2 is an abundant protein in human heart, it may play an important role in the regulation of cell differentiation and myofibrillogenesis of heart at defined subcellular compartment. Therefore, we hypothesized that FHL2 act as a multi-functional protein by the specific arrangement of the LIM domains of FHL2 and that one of the LIM domains of FHL2 can function as an anchor and localizes it into a specific subcellular compartment in a cell type specific manner to regulate myofibrillogenesis. From our results, we observed that FHL2 is localized at the focal adhesions of the C2C12, H9C2 myoblast as well as a nonmyogenic cell line, HepG2 cells. Colocalization of vinculin-CFP and FHL2-GFP at focal adhesions was also observed in cell lines. Site-directed mutagenesis, in turn, suggested that the second LIM domain-LIM2 is essential for its specific localization to focal adhesions. Moreover, FHL2 was observed along with F-actin and focal adhesion of C2C12 and H9C2 myotubes. Finally, we believe that FHL2 moves from focal adhesions and then stays at the Z-discs of terminally differentiated heart muscle.
authors: Department of Biochemistry, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China.
authors: H Y
authors: HY
authors: Li
authors: M
authors: M
authors: Kotaka
authors: S
authors: S
authors: Kostin
authors: S M
authors: SM
authors: Lee
authors: L D
authors: LD
authors: Kok
authors: K K
authors: KK
authors: Chan
authors: S K
authors: SK
authors: Tsui
authors: J
authors: J
authors: Schaper
authors: R
authors: R
authors: Zimmermann
authors: C Y
authors: CY
authors: Lee
authors: K P
authors: KP
authors: Fung
authors: M M
authors: MM
authors: Waye
mesh_terms: D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D002454:Cell Differentiation; D002460:Cell Line; D005136:Eye Proteins / Q000378:metabolism*; D022001:Focal Adhesions / Q000378:metabolism* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D060850:LIM-Homeodomain Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009124:Muscle Proteins; D009132:Muscles / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure*; D016297:Mutagenesis, Site-Directed; D009206:Myocardium / Q000378:metabolism / Q000648:ultrastructure*; D009210:Myofibrils / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016415:Sequence Alignment; D014157:Transcription Factors; D014407:Tumor Cells, Cultured; D016596:Vinculin / Q000378:metabolism
chemical_list: D000199:Actins; D005136:Eye Proteins; C105987:FHL2 protein, human; D018398:Homeodomain Proteins; D060850:LIM-Homeodomain Proteins; C044539:LIM2 protein, human; D008164:Luminescent Proteins; D008565:Membrane Proteins; D009124:Muscle Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D016596:Vinculin; D049452:Green Fluorescent Proteins