title: The interconversion of protein phosphatase 2A between PP2A1 and PP2A0 during retinoic acid-induced granulocytic differentiation and a modification on the catalytic subunit in S phase of HL-60 cells.
abstract: Alterations in protein phosphatase 2A (PP2A) during retinoic acid-induced differentiation of HL-60 cells have been investigated. PP2A activity of HL-60 cells for phosphorylated myelin basic protein showed a sharp and transient increase after 18-h treatment with 1 microM retinoic acid, which corresponded to G1/S boundary of the cell cycle. This PP2A of the 18-h treated cells was eluted from a DEAE-Sepharose column with 0.13 M NaCl, while PP2A from control cells was eluted with 0.23 M NaCl. The phosphorylase phosphatase activity of PP2A in the 0.13 M eluate was greatly enhanced in the presence of protamine compared with that of the later eluting PP2A. Immunoblot analyses with antisera against B' and B alpha subunits showed that the PP2A in the 0.13 M NaCl eluate from 18-h retinoic acid-treated cells was PP2A0 (AC-B'), whereas the PP2A eluted with 0.23 M NaCl from 24-h retinoic acid-treated cells and 0-, 18-, and 24-h control cells was PP2A1 (AC-B alpha). These results strongly suggest that PP2A undergoes a transient and reversible interconversion of holoenzyme forms during the initial stage of retinoic acid-induced granulocytic differentiation. PP2A activity assayed after dissociation of the catalytic subunit, for phosphorylase as substrate, showed a sharp and transient decrease in S phase of HL-60 cells irrespective of the presence or absence of retinoic acid. Immunoblot analyses with antisera against C-terminus and N-terminus of the catalytic subunit of PP2A suggested that a modification at the C-terminus is responsible for the decrease in PP2A activity. Immunoreactivity to the C-terminal antibody was restored after treatments of the S-phase extract with alkali or ethanol, the conditions which remove the methyl group from the C-terminus. These results suggest that the C-terminus of PP2A catalytic subunit is transiently methylated in S phase of HL-60 cells.
authors.affiliation: Section of Biochemistry, Hokkaido University, Sapporo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Zhu
authors.forename: S
authors.initials: S
authors.lastname: Matsuzawa
authors.forename: Y
authors.initials: Y
authors.lastname: Mizuno
authors.forename: C
authors.initials: C
authors.lastname: Kamibayashi
authors.forename: M C
authors.initials: MC
authors.lastname: Mumby
authors.forename: N
authors.initials: N
authors.lastname: Andjelkovic
authors.forename: B A
authors.initials: BA
authors.lastname: Hemmings
authors.forename: K
authors.initials: K
authors.lastname: Onoé
authors.forename: K
authors.initials: K
authors.lastname: Kikuchi
mesh_terms: D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division; D006098:Granulocytes / Q000166:cytology*; D018922:HL-60 Cells / Q000166:cytology / Q000201:enzymology*; D006801:Humans; D046911:Macromolecular Substances; D008745:Methylation; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D054648:Protein Phosphatase 2; D016196:S Phase; D014212:Tretinoin / Q000494:pharmacology*
chemical_list: D046911:Macromolecular Substances; D014212:Tretinoin; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2