Dissociation of FAK/p130CAS/c-Src Complex during Mitosis: Role of Mitosis-specific Serine Phosphorylation of FAK

Yamakita, Yoshihiko; Totsukawa, Go; Fry, David W.; Zhang, Xiaoe; Hanks, Steven K.; Matsumura, Fumio

doi:10.1083/jcb.144.2.315

Cited by 115 publications

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References 75 publications

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“…Several reports pointed out a tyrosine dephosphorylation of the components of the focal adhesion complex including FAK, paxillin and p130 Cas associated with ser/thr phosphorylation of these proteins and the dissociation of the complex during the M phase of the cell cycle. This suggests that focal adhesion protein interaction is linked to ser/thr phosphorylation status [23,24]. Aurora-A is a member of Aurora kinase family that includes three members, Aurora-A, -B and -C. All of them are regulators of mitosis and Aurora-A and -B are overexpressed in a variety of tumor cell lines suggesting a role in tumorigenesis [25].…”

Section: Introductionmentioning

confidence: 99%

Phosphorylation of human enhancer of filamentation (HEF1) on serine 369 induces its proteasomal degradation

Hivert

Pierre

Raingeaud

2009

Biochemical Pharmacology

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Please cite this article as: Hivert V, Pierre J, Raingeaud J, Phosphorylation of human enhancer of filamentation (HEF1) on serine 369 induces its proteasomal degradation, Biochemical Pharmacology (2008Pharmacology ( ), doi:10.1016Pharmacology ( /j.bcp.2009 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Cas family. HEF1 is present at focal adhesions and is involved in integrin signalling mediating cytoskeleton reorganization associated with cell migration, adhesion or apoptosis.HEF1 functions are regulated in part by phosphorylation on tyrosine residues. HEF1 is also phosphorylated on serines/threonines leading to two isoforms refered to as p105 and p115. In most cases, the serine/threonine kinase(s) responsible for HEF1 phosphorylation have not been identified. In the present study, we have investigated HEF1 ser/thr phosphorylation. In the HCT-116 cell line transiently overexpressing Flag-HEF1 we showed that Hesperadin, a synthetic indolinone displaying antiproliferative effect and described as an inhibitor of various kinases including Aurora-B, prevented HEF1 phosphorylation induced by the ser/thr phosphatase PP2A inhibitor : okadaic acid (OA). In addition we showed that conversion of endogenous HEF1 p105 to p115 in HaCaT cells was prevented upon treatment with Hesperadin, resulting in accumulation of p105HEF1. We also identified serine 369 as the target site of phosphorylation by this Hesperadin-inhibited kinase in HCT-116. Finally, we * ManuscriptPage 2 of 37 A c c e p t e d M a n u s c r i p t 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 provide evidence that phosphorylation on serine 369 but not phosphorylation on serine 296, triggers HEF1 degradation by the proteasomal machinery. These data suggest that conversion of p105 to p115 results from a ser-369-dependent phosphorylation mediated by an Hesperadin-sensitive kinase and regulates the half-life of HEF1.Keywords: human enhancer of filamentation 1, protein phosphatase 2A, Hesperadin, protein stability.

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Section: Introductionmentioning

confidence: 99%

Phosphorylation of human enhancer of filamentation (HEF1) on serine 369 induces its proteasomal degradation

Hivert

Pierre

Raingeaud

2009

Biochemical Pharmacology

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“…More recently, it has become clear that FAK is also phosphorylated at multiple serine residues [32][33][34][35][36][37]. The proximity of these phosphorylated serine residues to sites at which FAK interacts with other proteins suggests a possible function in the regulation of the assembly of FAK signaling complexes [6].…”

Section: Introductionmentioning

confidence: 99%

Differential FAK phosphorylation at Ser-910, Ser-843 and Tyr-397 induced by angiotensin II, LPA and EGF in intestinal epithelial cells

Jiang

Sinnett‐Smith

2007

Cellular Signalling

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A rapid increase in the tyrosine phosphorylation of the non-receptor tyrosine kinase FAK is a prominent early event in fibroblasts stimulated by a variety of signaling molecules. However, a variety of epithelial cells, including intestinal epithelial cells, show a high basal level of tyrosine phosphorylated FAK that is only slightly further increased by addition of G protein-coupled receptors (GPCR) agonists or growth factors. In this study, we determined whether these stimuli could elicit FAK phosphorylation at serine residues, including Ser-910 and Ser-843. Our results show that multiple agonists including angiotensin II (ANGII), lysophosphatidic acid (LPA), phorbol esters and EGF induced a striking stimulation of FAK phosphorylation at Ser-910 in rat intestinal epithelial IEC-18 cells via an ERK-dependent pathway. In striking contrast, none of these stimuli promoted a significant further increase in FAK phosphorylation at Tyr-397 in these cells. These results were extended using cultures of polarized human colonic epithelial T84 cells. We found that either carbachol or EGF promoted a striking ERK-dependent phosphorylation of FAK at Ser-910, but these agonists caused only slight stimulation of FAK at Tyr-397 in T84 cells. In addition, we demonstrated that GPCR agonists also induced a dramatic increase of FAK phosphorylation at Ser-843 in either IEC-18 or T84 cells. Our results indicate that Ser-910 and Ser-843, rather than Tyr-397, are prominent sites differentially phosphorylated in response to neurotransmitters, bioactive lipids, tumor promoters and growth factors in intestinal epithelial cells.

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“…However, because adhesion is most readily observable in interphase cells, and focal adhesions tend to minimize or disappear in mitotic cells, accompanied by the down-regulation of some key focal adhesion-associated signaling proteins (Yamaguchi et al, 1997;Law et al, 1998;Yamakita et al, 1999), little work has investigated a possible requirement or alternative roles for adhesion-associated proteins in M phase. Similarly, it is only in the past several years that investigations of the nuclear cell cycle have documented the coordination of signaling events in time and space during M-phase progression, emphasizing the roles of proteins associated with structures such as the centrosome, the contractile ring, the central spindle, and the midbody (Raff et al, This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E05-03-0237) on January 4, 2006. 2002; reviewed in Glotzer, 2001).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, a number of proteins most studied in the context of action at focal adhesions, including zyxin (Hirota et al, 2000), paxillin (Yamaguchi et al, 1997), FAK (Yamakita et al, 1999;Fresu et al, 2001;Ma et al, 2001), and the Cas family member HEF1 (Law et al, 1998), have been implicated in mitotic control, based on dramatic changes in their phosphorylation, or their localization to mitotic structures, in M phase. We have begun to investigate the role of HEF1 as a direct regulator of mitotic progression, and through this work, we have recently demonstrated that HEF1 associates with and positively regulates the Aurora-A kinase at mitotic entry (Pugacheva and Golemis, 2005).…”

Section: Introductionmentioning

confidence: 99%

Deregulation of HEF1 Impairs M-Phase Progression by Disrupting the RhoA Activation Cycle

Dadke

Jarnik

Pugacheva

et al. 2006

MBoC

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The focal adhesion-associated signaling protein HEF1 undergoes a striking relocalization to the spindle at mitosis, but a function for HEF1 in mitotic signaling has not been demonstrated. We here report that overexpression of HEF1 leads to failure of cells to progress through cytokinesis, whereas depletion of HEF1 by small interfering RNA (siRNA) leads to defects earlier in M phase before cleavage furrow formation. These defects can be explained mechanistically by our determination that HEF1 regulates the activation cycle of RhoA. Inactivation of RhoA has long been known to be required for cytokinesis, whereas it has recently been determined that activation of RhoA at the entry to M phase is required for cellular rounding. We find that increased HEF1 sustains RhoA activation, whereas depleted HEF1 by siRNA reduces RhoA activation. Furthermore, we demonstrate that chemical inhibition of RhoA is sufficient to reverse HEF1-dependent cellular arrest at cytokinesis. Finally, we demonstrate that HEF1 associates with the RhoA-GTP exchange factor ECT2, an orthologue of the Drosophila cytokinetic regulator Pebble, providing a direct means for HEF1 control of RhoA. We conclude that HEF1 is a novel component of the cell division control machinery and that HEF1 activity impacts division as well as cell attachment signaling events. INTRODUCTIONAs points of structural linkage between the extracellular matrix (ECM) and the intracellular cytoskeleton, focal adhesions possess a complex function. For example, during migration, cells must rapidly break down and reform adhesions with the ECM, providing force for propulsion (Lauffenburger and Horwitz, 1996). At mitotic entry, cultured cells round up and decrease adhesion to the ECM; at mitotic exit, basal attachments reassemble and contribute to the force generation required for efficient progress through cytokinesis and reentry into G 1 . In interphase cells, the formation of novel focal adhesion-ECM interactions can specify cellular differentiation by activating specific signaling cascades culminating in the induction of differentiationpromoting transcription factors, and in parallel enforce removal from the cell cycle (Boudreau and Bissell, 1998). In many cell types, sustained loss of adhesion is a sufficient stimulus to induce apoptosis (anoikis) (Frisch and Francis, 1994), a surveillance mechanism against cancer, inhibiting the formation of micrometastases. Hence, one frequent effect of oncogenic transformation is the circumvention of the adhesion-viability coupling, leading to acquisition by cancer cells of the ability to grow in an anchorage-independent manner (Schwartz, 1997). Based on these diverse biological roles, there has been considerable research effort directed at elucidating the signaling role of focal adhesion-associated proteins (Schlaepfer et al., 1999).HEF1, p130Cas, and Efs/Sin define the Cas family of proteins Bouton et al., 2001). In interphase cells, Cas proteins predominantly localize to focal adhesions. During initial integrin engagement, induced by cell a...

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Dissociation of FAK/p130CAS/c-Src Complex during Mitosis: Role of Mitosis-specific Serine Phosphorylation of FAK

Cited by 115 publications

References 75 publications

Phosphorylation of human enhancer of filamentation (HEF1) on serine 369 induces its proteasomal degradation

Phosphorylation of human enhancer of filamentation (HEF1) on serine 369 induces its proteasomal degradation

Differential FAK phosphorylation at Ser-910, Ser-843 and Tyr-397 induced by angiotensin II, LPA and EGF in intestinal epithelial cells

Deregulation of HEF1 Impairs M-Phase Progression by Disrupting the RhoA Activation Cycle

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