Leucine Carboxyl Methyltransferase 1 (LCMT-1) Methylates Protein Phosphatase 4 (PP4) and Protein Phosphatase 6 (PP6) and Differentially Regulates the Stable Formation of Different PP4 Holoenzymes

J Biol Chem. 2016 Sep 30;291(40):21008-21019. doi: 10.1074/jbc.M116.739920. Epub 2016 Aug 9.


The protein phosphatase 2A (PP2A) subfamily of phosphatases, PP2A, PP4, and PP6, are multifunctional serine/threonine protein phosphatases involved in many cellular processes. Carboxyl methylation of the PP2A catalytic subunit (PP2Ac) C-terminal leucine is regulated by the opposing activities of leucine carboxyl methyltransferase 1 (LCMT-1) and protein phosphatase methylesterase 1 (PME-1) and regulates PP2A holoenzyme formation. The site of methylation on PP2Ac is conserved in the catalytic subunits of PP4 and PP6, and PP4 is also methylated on that site, but the identities of the methyltransferase enzyme for PP4 are not known. Whether PP6 is methylated is also not known. Here we use antibodies specific for the unmethylated phosphatases to show that PP6 is carboxyl-methylated and that LCMT-1 is the major methyltransferase for PP2A, PP4, and PP6 in mouse embryonic fibroblasts (MEFs). Analysis of PP2A and PP4 complexes by blue native polyacrylamide gel electrophoresis (BN-PAGE) indicates that PP4 holoenzyme complexes, like those of PP2A, are differentially regulated by LCMT-1, with the PP4 regulatory subunit 1 (PP4R1)-containing PP4 complex being the most dramatically affected by the LCMT-1 loss. MEFs derived from LCMT-1 knock-out mouse embryos have reduced levels of PP2A B regulatory subunit and PP4R1 relative to control MEFs, indicating that LCMT-1 is important for maintaining normal levels of these subunits. Finally, LCMT-1 homozygous knock-out MEFs exhibited hyperphosphorylation of HDAC3, a reported target of the methylation-dependent PP4R1-PP4c complex. Collectively, our data suggest that LCMT-1 coordinately regulates the carboxyl methylation of PP2A-related phosphatases and, consequently, their holoenzyme assembly and function.

Keywords: Lcmt-1; PP4; PP6; methyltransferase; protein complex; protein methylation; protein phosphatase 2 (PP2A); protein phosphorylation; signaling.

MeSH terms

  • Animals
  • Cells, Cultured
  • Embryo, Mammalian / enzymology*
  • Fibroblasts / enzymology*
  • Histone Deacetylases / genetics
  • Histone Deacetylases / metabolism
  • Holoenzymes / genetics
  • Holoenzymes / metabolism
  • Methylation
  • Mice
  • Mice, Knockout
  • Phosphoprotein Phosphatases / genetics
  • Phosphoprotein Phosphatases / metabolism*
  • Phosphorylation / genetics
  • Protein O-Methyltransferase / genetics
  • Protein O-Methyltransferase / metabolism*


  • Holoenzymes
  • Lcmt1 protein, mouse
  • Protein O-Methyltransferase
  • Phosphoprotein Phosphatases
  • protein phosphatase 4
  • protein phosphatase 6
  • Histone Deacetylases
  • histone deacetylase 3