Protein phosphatase 2A is targeted to cell division control protein 6 by a calcium-binding regulatory subunit.
The cell division control protein 6 (Cdc6) is essential for formation of pre-replication complexes at origins of DNA replication. Phosphorylation of Cdc6 by cyclin-dependent kinases inhibits ubiquitination of Cdc6 by APC/C(cdh1) and degradation by the proteasome. Experiments described here show that the PR70 member of the PPP2R3 family of regulatory subunits targets protein phosphatase 2A (PP2A) to Cdc6. Interaction with Cdc6 is mediated by residues within the C terminus of PR70, whereas interaction with PP2A requires N-terminal sequences conserved within the PPP2R3 family. Two functional EF-hand calcium-binding motifs mediate a calcium-enhanced interaction of PR70 with PP2A. Calcium has no effect on the interaction of PR70 with Cdc6 but enhances the association of PP2A with Cdc6 through its effects on PR70. Knockdown of PR70 by RNA interference results in an accumulation of endogenous and expressed Cdc6 protein that is dependent on the cyclin-dependent protein kinase phosphorylation sites on Cdc6. Knockdown of PR70 also causes G(1) arrest, suggesting that PR70 function is critical for progression into S phase. These observations indicate that PP2A can be targeted in a calcium-regulated manner to Cdc6 via the PR70 subunit, where it plays a role in regulating protein phosphorylation and stability.
Davis
Anthony J
AJ
Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, Texas 75390-9041, USA.
Yan
Zhen
Z
Martinez
Bobbie
B
Mumby
Marc C
MC
D020816:Amino Acid Motifs / Q000502:physiology; D000818:Animals; D015703:Antigens, CD; D019556:COS Cells; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004261:DNA Replication / Q000502:physiology; D016193:G1 Phase / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D018741:Replication Origin / Q000502:physiology; D016196:S Phase / Q000502:physiology*; D054875:Ubiquitination / Q000502:physiology*
D015703:Antigens, CD; C103706:CDC6 protein, human; C513384:CDH1 protein, human; D015820:Cadherins; D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D021122:Protein Subunits; D054648:Protein Phosphatase 2; D046988:Proteasome Endopeptidase Complex; D002118:Calcium