Subcellular trafficking of exogenously expressed interferon-beta in Madin-Darby canine kidney cells.
We have recently demonstrated that when IFN-beta was exogenously expressed in epithelial cells, transiently expressed IFN-beta was predominantly secreted from the cell side to which the transfection was performed, while stably expressed one was almost equally secreted to the apical and basolateral sides. In the present study, we analyzed the subcellular transport of IFN-beta using confocal imaging with green fluorescent protein (GFP)-tagged IFN-beta in Madin-Darby canine kidney (MDCK) cells. Stably expressed and transiently expressed human IFN-beta (HuIFN-beta)-GFPs were seen in upper regions of the nucleus. In stable HuIFN beta-GFP-producing transformants, transiently expressed mouse IFN-beta (MuIFN-beta) was apparently co-localized with the bulk of the constitutive HuIFN beta-GFP proteins at TGN, and a significant quantity of them then appeared to pass into distinct post-TGN vesicles, accepting either type of IFN. Meanwhile, when cells were co-transfected with both expression vectors, transiently expressed both IFNs tended to co-localize not only at TGN but in post-TGN vesicles. These results suggest that stably and transiently expressed IFN-betas, albeit co-localized at TGN, were transported through apparently discriminated post-TGN routes.
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
Masato
M
Maruyama
Teruko
T
Nishio
Takako
T
Kato
Toyokazu
T
Yoshida
Chisaki
C
Ishida
Yoshihiko
Y
Watanabe
Makiya
M
Nishikawa
Yasufumi
Y
Kaneda
Yoshinobu
Y
Takakura
D000818:Animals; D002460:Cell Line; D016764:Cell Polarity / Q000502:physiology*; D004285:Dogs; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D016899:Interferon-beta / Q000235:genetics* / Q000378:metabolism*; D007668:Kidney / Q000166:cytology*; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D021381:Protein Transport / Q000502:physiology*; D021601:trans-Golgi Network / Q000502:physiology*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D016899:Interferon-beta