Molecular Dissection of Internalization of Porphyromonas gingivalis by Cells using Fluorescent Beads Coated with Bacterial Membrane Vesicle

Cell Structure and Function

Ichiro Nakagawa⁴  Atsuo Amano²  Kyohei Umebayashi³  Yoshinobu Nakanishi¹  Kayoko Tsuda¹  Tamotsu Yoshimori³  Hiroaki Inaba² 

[1] Graduate School of Medical Science, Kanazawa University;Department of Oral Frontier Biology, Osaka University Graduate School of Dentistry;Department of Cell Genetics, National Institute of Genetics/SOKENDAI;Department of Oral and Molecular Microbiology, Osaka University Graduate School of Dentistry

关键词: P. gingivalis;    membrane vesicles;    dynamin;    actin;    microtubule;    lipid raft;   

DOI  :  10.1247/csf.30.81

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(53)Cited-By(17)Porphyromonas gingivalis is one of the causative agents of adult periodontitis, and has been reported to be internalized by nonphagocytic epithelial cells. However, the mechanism for the internalization remains unclear. In the present study, we addressed this issue using fluorescent beads coated with bacterial membrane vesicles (MVs) that retain surface components of P. gingivalis. We established an assay system in which we could easily quantify the bead internalization to cells. MVs-coated beads were internalized by HeLa cells in kinetics similar to that of living bacteria. The internalization depended on dynamin but not clathrin. The beads were internalized through the actin-mediated pathway that is controlled by phosphatidylinositol (PI) 3-kinase. The dynamics of microtubule assembly and disassembly was also required. Further, the treatment of cells with cholesterol-binding reagents significantly inhibited bead internalization, and the internalized beads were apparently colocalized with ganglioside GM1 and caveolin-1, which suggest the involvement of the lipid raft in the process. These results suggest that P. gingivalis accomplishes its internalization utilizing membrane lipid raft and cytoskeletal functions of the target cells.

【 授权许可】

Unknown