Generation of active protein phosphatase 2A is coupled to holoenzyme assembly.
Protein phosphatase 2A (PP2A) is a prime example of the multisubunit architecture of protein serine/threonine phosphatases. Until substrate-specific PP2A holoenzymes assemble, a constitutively active, but nonspecific, catalytic C subunit would constitute a risk to the cell. While it has been assumed that the severe proliferation impairment of yeast lacking the structural PP2A subunit, TPD3, is due to the unrestricted activity of the C subunit, we recently obtained evidence for the existence of the C subunit in a low-activity conformation that requires the RRD/PTPA proteins for the switch into the active conformation. To study whether and how maturation of the C subunit is coupled with holoenzyme assembly, we analyzed PP2A biogenesis in yeast. Here we show that the generation of the catalytically active C subunit depends on the physical and functional interaction between RRD2 and the structural subunit, TPD3. The phenotype of the tpd3Delta strain is therefore caused by impaired, rather than increased, PP2A activity. TPD3/RRD2-dependent C subunit maturation is under the surveillance of the PP2A methylesterase, PPE1, which upon malfunction of PP2A biogenesis, prevents premature generation of the active C subunit and holoenzyme assembly by counteracting the untimely methylation of the C subunit. We propose a novel model of PP2A biogenesis in which a tightly controlled activation cascade protects cells from untargeted activity of the free catalytic PP2A subunit.
Hombauer
Hans
H
Department of Medical Biochemistry, Max F. Perutz Laboratories, Medical University of Vienna, Vienna, Austria.
Weismann
David
D
Mudrak
Ingrid
I
Stanzel
Claudia
C
Fellner
Thomas
T
Lackner
Daniel H
DH
Ogris
Egon
E
D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism; D004789:Enzyme Activation / Q000502:physiology; D020035:Holoenzymes / Q000378:metabolism; D008745:Methylation; D008954:Models, Biological; D010749:Phosphoprotein Phosphatases / Q000096:biosynthesis / Q000378:metabolism*; D054648:Protein Phosphatase 2; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D013379:Substrate Specificity
C071246:CDC55 protein, S cerevisiae; D018797:Cell Cycle Proteins; D020035:Holoenzymes; C502933:RRD2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D002265:Carboxylic Ester Hydrolases; C119357:protein phosphatase methylesterase-1; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2