[Actin organization regulated by lipids].
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Biomedical subjects
Publications and source records attributed to Takuma Kishimoto.
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Specific changes in membrane lipid composition are implicated in actin cytoskeletal organization, vesicle formation, and control of cell polarity. Cdc50p, a membrane protein in the endosomal/trans-Golgi network compartments, is a noncatalytic subunit of Drs2p, which is implicated in translocation of phospholipids across lipid bilayers. We found that the cdc50Delta mutation is synthetically lethal with mutations affecting the late steps of ergosterol synthesis (erg2 to erg6). Defects in cell polarity and actin organization were observed in the cdc50Delta erg3Delta mutant. In particular, actin patches, which are normally found at cortical sites, were assembled intracellularly along with their assembly factors, including Las17p, Abp1p, and Sla2p. The exocytic SNARE Snc1p, which is recycled by an endocytic route, was also intracellularly accumulated, and inhibition of endocytic internalization suppressed the cytoplasmic accumulation of both Las17p and Snc1p. Simultaneous loss of both phospholipid asymmetry and sterol structural integrity could lead to accumulation of endocytic intermediates capable of initiating assembly of actin patches in the cytoplasm.
The influence of gamma-ray irradiation on a cementum-impregnated gelatine membrane (CGM) was analyzed with emphasis on its function during periodontal regeneration. In brief, proteins were extracted from gamma-ray irradiated cementum (gammaC). With the gammaC protein, sample cells (gingival fibroblasts, periodontal ligament cells, and alveolar bone cells) were co-cultured, and cytological parameters (cell attachment, cell differentiation and alkaline phosphatase activity) were analyzed. Additionally, kinetics of some gene expression was analysed using reverse transcript RT-PCR, which included osteoproteogerin (OPG)/osteoclastogenesis inhibitory factor (OCIF) mRNA. BMP-2 and osteonectin were resistant to gamma-rays, and other cytokines involved in regeneration were decreased. Thus, the attachment activity of osteoblasts to gammaC protein was higher than that of non-irradiated cementum (control C). The expression of OPG/OCIF mRNA was lower in co-cultured cells with gammaC protein than those with in control C protein. Together the results imply that some cytokine in intact cementum prevents the attachment (differentiation) of bone cells onto the root surface, which may explain why the introduction of CGM following gingival flap surgery induces new cementum, new ligament and new bone formation, but CGM irradiated with gamma-rays for clinical use causes ankylosis.