Cell adhesion regulates Ser/Thr phosphorylation and proteasomal degradation of HEF1.
Human enhancer of filamentation 1 (HEF1), a multifunctional docking protein of the Cas family, participates in integrin and growth factor signaling pathways that regulate global cellular processes including growth, motility and apoptosis. HEF1 consists of two isoforms, p105 and p115, the larger molecular weight form resulting from Ser/Thr phosphorylation of p105HEF1. The molecular mechanisms that regulate the interconversion of the two HEF1 species as well as the function of HEF1 Ser/Thr phosphorylation are unknown. Our study reveals that cell adhesion and detachment regulate the interconversion of the two HEF1 isoforms. Experiments using various inhibitors of cytoskeletal organization indicated that disruption of actin microfilaments but not intermediate filaments or microtubules resulted in a complete conversion of p115HEF1 to p105HEF1. The conversion of p115HEF1 to p105HEF1 was prevented by inhibition of protein phosphatase 2A (PP2A), suggesting that cytoskeletal regulation of PP2A activity controlled the dephosphorylation of p115HEF1. Degradation of endogenous HEF1 was dependent on proteasomes with the p115 species of HEF1 being preferentially targeted for turnover. Dephosphorylation of HEF1 by suspending cells or disrupting actin filaments protected HEF1 from degradation. These results suggest that the adhesion-dependent actin organization regulates proteasomal turnover of HEF1 through the activity of PP2A.
Zheng
Mingzhe
M
Center for Cell Biology and Cancer Research, Albany Medical College, Albany, NY 12208, USA.
McKeown-Longo
Paula J
PJ
D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D002460:Cell Line; D003078:Colchicine / Q000378:metabolism; D015638:Cytochalasin D / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism; D008387:Marine Toxins; D008870:Microtubules / Q000378:metabolism; D015739:Nocodazole; D019384:Nucleic Acid Synthesis Inhibitors / Q000378:metabolism; D019319:Okadaic Acid / Q000378:metabolism; D010080:Oxazoles / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D054648:Protein Phosphatase 2; D011714:Pyrans / Q000378:metabolism; D012694:Serine / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D013141:Spiro Compounds / Q000378:metabolism; D013912:Threonine / Q000378:metabolism*; D050257:Tubulin Modulators / Q000378:metabolism
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D008387:Marine Toxins; C099942:NEDD9 protein, human; D019384:Nucleic Acid Synthesis Inhibitors; D010080:Oxazoles; D010750:Phosphoproteins; D061988:Proteasome Inhibitors; D020033:Protein Isoforms; D011714:Pyrans; D013141:Spiro Compounds; D050257:Tubulin Modulators; C053079:tautomycin; D019319:Okadaic Acid; D015638:Cytochalasin D; D013912:Threonine; D012694:Serine; C059041:calyculin A; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D046988:Proteasome Endopeptidase Complex; D015739:Nocodazole; D003078:Colchicine