Targeting of FAK Ser910 by ERK5 and PP1delta in non-stimulated and phorbol ester-stimulated cells.
Ser910 of FAK (focal adhesion kinase) was phosphorylated in fibroblasts treated with the phorbol ester PMA and dephosphorylated by PP1d (protein phosphatase 1d), as indicated by shRNA (small-hairpin RNA) gene silencing. Ser910 of FAK was reported previously to be an ERK (extracellular-signal-regulated kinase) 1/2 target in cells treated with phorbol esters. In contrast, various approaches, including the use of the MEK (mitogen-activated protein kinase/ERK kinase) inhibitors UO126 and CI-1040 to inhibit ERK1/2 pointed to the involvement of ERK5. This hypothesis was confirmed by: (i) shRNA ERK5 gene silencing, which resulted in complete pSer910 loss in non-stimulated and PMA-stimulated cells; (ii) direct phosphorylation of recombinant FAK by ERK5; and (iii) ERK5 activation by PMA. PMA stimulation and ERK5 silencing in MDA-MB 231 and MDA-MB 361 breast cancer cells indicated Ser910 targeting by ERK5 also in these cells. Given the proximity of Ser910 to the FAT (focal adhesion targeting) regulatory domain of FAK, cell proliferation and morphology were investigated in FAK-/- cells expressing S910A mutant FAK. The cell growth rate decreased and exposure to PMA induced peculiar morphological changes in cells expressing S910A, with respect to wild-type FAK, suggesting a role for Ser910 in these processes. The present study indicates, for the first time, the phosphorylation of Ser910 of FAK by ERK5 and its dephosphorylation by PP1d, and suggested a role for Ser910 in the control of cell shape and proliferation.
Villa-Moruzzi
Emma
E
Dipartimento di Patologia Sperimentale, Sezione Patologia Generale, via Roma 55, 56126 Pisa, Italy. villa@biomed.unipi.it
D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D004789:Enzyme Activation / Q000187:drug effects; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D020868:Gene Silencing; D006801:Humans; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D048054:Mitogen-Activated Protein Kinase 7 / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D010703:Phorbol Esters / Q000494:pharmacology*; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D012694:Serine / Q000235:genetics / Q000378:metabolism
D010703:Phorbol Esters; D012694:Serine; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048054:Mitogen-Activated Protein Kinase 7; C524311:PP1delta protein, rat; D010749:Phosphoprotein Phosphatases
FEBS Lett. 2001 Jul 27;502(1-2):21-4
11478941
Methods Enzymol. 1991;201:21-7
1719341
Mol Cell Biol. 2002 Jan;22(1):270-85
11739740
Int J Cancer. 2002 Jul 20;100(3):276-82
12115541
J Cell Sci. 2003 Apr 15;116(Pt 8):1409-16
12640026
Front Biosci. 2003 May 1;8:d982-96
12700132
J Biol Chem. 2003 Jun 20;278(25):22631-43
12692126
Curr Biol. 2003 Nov 11;13(22):R886-8
14614847
Physiol Rev. 2004 Jan;84(1):1-39
14715909
Methods Mol Biol. 2004;250:113-26
14755084
Nat Cell Biol. 2004 Feb;6(2):154-61
14743221
J Mammary Gland Biol Neoplasia. 2003 Oct;8(4):395-408
14985636
Trends Cell Biol. 2004 May;14(5):241-9
15130580
J Cell Physiol. 2004 Aug;200(2):213-22
15174091
Biochim Biophys Acta. 2004 Jul 5;1692(2-3):77-102
15246681
Physiol Rev. 2004 Oct;84(4):1315-39
15383653
Int J Oncol. 2004 Nov;25(5):1383-8
15492829
Eur J Biochem. 1983 Jun 15;133(2):455-61
6303789
Biochem J. 1997 May 15;324 ( Pt 1):141-9
9164850
Mol Biol Cell. 1997 Apr;8(4):663-73
9247646
Cell Signal. 1998 Apr;10(4):225-32
9617479
Anticancer Res. 2000 Jul-Aug;20(4):2265-71
10953284
Mol Biol Cell. 2001 Jan;12(1):1-12
11160818
Methods Mol Biol. 1998;93:35-40
9664525
Cell Adhes Commun. 1998 Jun;5(4):297-305
9762470
J Cell Biol. 1999 Jan 25;144(2):315-24
9922457
Mol Cell Biol. 1999 Jul;19(7):4806-18
10373530
J Cell Sci. 2005 Apr 15;118(Pt 8):1663-71
15797923
J Biol Chem. 2005 Jun 24;280(25):24212-20
15845548
Mol Cell Biochem. 2005 Apr;272(1-2):85-90
16010975
Mol Cell Biol. 2005 Oct;25(19):8553-66
16166637
Biochem J. 2005 Oct 15;391(Pt 2):359-70
15975092
J Cell Sci. 2005 Nov 1;118(Pt 21):4931-5
16254240
Cell Signal. 2006 Jun;18(6):753-60
16376520
EMBO Rep. 2006 Aug;7(8):782-6
16880823
Curr Opin Cell Biol. 2006 Oct;18(5):516-23
16919435
Biochem J. 2001 Sep 1;358(Pt 2):407-14
11513739