Localization of the PP2A B56gamma regulatory subunit at the Golgi complex: possible role in vesicle transport and migration.
The BL6 subline was derived from the F10 line, which was derived from the B16 mouse melanoma cell line. BL6 cells are more invasive than F10 cells and differ genetically from F10 cells by an alteration of the gene encoding the B56gamma regulatory subunit of protein phosphatase 2A (PP2A). This alteration results in the transcription of mRNA encoding a truncated variant of the B56gamma1 isoform (Deltagamma1). When F10 cells were stained with a polyclonal antibody that recognizes three B56gamma isoforms, B56gamma1, B56gamma2, and B56gamma3, the immunofluorescent signals co-localized well with the cis-Golgi marker proteins. When BL6 cells were fractionated in a sucrose gradient, B56gamma1 and B56gamma2, but not B56gamma3, were present in the Golgi-enriched fraction. This fraction also contained the catalytic subunit of PP2A. FLAG-tagged Deltagamma1 preferentially localized to the trans-Golgi area rather than the cis-Golgi. This localization was the same as that of FLAG-tagged B56gamma1. NIH3T3 cells stably expressing Deltagamma1 transported a mutant viral protein from the endoplasmic reticulum to the plasma membrane much faster than wild-type cells. Their directional migration, as assessed by the advance of cells into a cell-free area, was also elevated. As Deltagamma1 reduces the activity of the B56gamma-containing PP2A holoenzymes, these results suggest that the normal holoenzymes suppress vesicle transport and that Deltagamma1 might increase the invasive ability of BL6 cells by activating Golgi function.
Ito
Akihiko
A
Department of Pathology, Osaka University Medical School/Graduate School of Frontier Bioscience, Suita, Osaka, Japan. aito@patho.med.osaka-u.ac.jp
Koma
Yu-ichiro
Y
Sohda
Miwa
M
Watabe
Kenji
K
Nagano
Teruaki
T
Misumi
Yoshio
Y
Nojima
Hiroshi
H
Kitamura
Yukihiko
Y
D016475:3T3 Cells; D000818:Animals; D002453:Cell Cycle; D002458:Cell Fractionation / Q000379:methods; D017931:DNA Primers; D006056:Golgi Apparatus / Q000201:enzymology*; D008546:Melanoma, Experimental / Q000201:enzymology* / Q000473:pathology*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids; D054648:Protein Phosphatase 2; D021122:Protein Subunits / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection
D017931:DNA Primers; D021122:Protein Subunits; D011994:Recombinant Proteins; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2