Aurora-B associated protein phosphatases as negative regulators of kinase activation

Sugiyama, Keiichi; Sugiura, Kazumitsu; Hara, Tomohiro; Sugimoto, Kenji; Shima, Hiroshi; Honda, Kanako; Furukawa, Koichi; Yamashita, Shunichi; Urano, Takeshi

doi:10.1038/sj.onc.1205432

Cited by 139 publications

(116 citation statements)

References 42 publications

Supporting

Mentioning

110

Contrasting

Order By: Relevance

“…4A). A role of PP1 and/or PP2A in regulating Aurora kinases has been described previously [29,30]. Autophosphorylation of a threonine residue in the kinase domain of Aurora is necessary to activate the kinases [29,34].…”

Section: Discussionmentioning

confidence: 99%

“…The above experiment showed that OA treatment revealed at least two different states of phosphorylation of HEF1. Moreover, OA has also been described as an activator of Aurora kinases A and B [29,30]. To examine the involvement of the Aurora kinases in HEF1 phosphorylation, we transiently transfected HCT-116 cells with 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 phosphorylation.…”

Section: Hef1 Is Phosphorylated In Vivo By An Hesperadin-sensitive Kimentioning

confidence: 99%

“…HEF1 interaction with Aurora-A leads to activation of the kinase while phosphorylated HEF1 fate is still unclear [22]. Aurora-B binds to and is negatively regulated by both PP1 and PP2A and is therefore activated by okadaic acid (OA), an inhibitor of these phosphatases [30]. Hesperadin, described as an inhibitor of Aurora-B kinase is a synthetic indolinone which displays antiproliferative properties and has entered phase I clinical trials as anti-cancer drug.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Hivert

Pierre

Raingeaud

2009

Biochemical Pharmacology

View full text Add to dashboard Cite

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.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Hef1 Is Phosphorylated In Vivo By An Hesperadin-sensitive Kimentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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

Hivert

Pierre

Raingeaud

2009

Biochemical Pharmacology

View full text Add to dashboard Cite

show abstract

“…The PP1 consensus binding site has been thoroughly characterized in plethora of proteins and was accurately defined (Egloff et al, 1997). Presence of two PP1 docking motifs was identified in various PP1 binding proteins (Ayllon et al, 2002;Bennett and Alphey, 2002;Sugiyama et al, 2002;Garcia et al, 2003). Analysis of the murine and human Fer amino-acid sequence revealed the presence of two PP1 docking motifs in the kinase domain of the two kinases and these motifs are separated by 42 aa from each other ( Figure 7a).…”

Section: Pp1a Associates With Fer and Is Downregulated In Fer-overexpmentioning

confidence: 99%

Downregulation of Fer induces PP1 activation and cell-cycle arrest in malignant cells

et al. 2006

View full text Add to dashboard Cite

Fer is a nuclear and cytoplasmic intracellular tyrosine kinase. Herein we show that Fer is required for cell-cycle progression in malignant cells. Decreasing the level of Fer using the RNA interference (RNAi) approach impeded the proliferation of prostate and breast carcinoma cells and led to their arrest at the G0/G1 phase. At the molecular level, knockdown of Fer resulted in the activation of the retinoblastoma protein (pRB), and this was reflected by profound hypo-phosphorylation of pRB on both cyclindependent kinase CDK4 and CDK2 phosphorylation sites. Dephosphorylation of pRB was not seen upon the direct targeting of either CDK4 or CDK2 expression, and was only partially achieved by the simultaneous depletion of these two kinases. Amino-acid sequence analysis revealed two protein phosphatase 1 (PP1) binding motifs in the kinase domain of Fer and the association of Fer with the pRB phosphatase PP1a was verified using co-immunoprecipitation analysis. Downregulation of Fer potentiated the activation of PP1a and overexpression of Fer decreased the enzymatic activity of that phosphatase. Our findings portray Fer as a regulator of cell-cycle progression in malignant cells and as a potential target for cancer intervention.

show abstract

“…This conclusion is corroborated by the stronger association of the human chromatin-associated phosphatase PP1-␦ to acetylated mitotic chromosomes (Figure 4). Indeed, a modified chromatin conformation may stimulate phosphatase loading on H3 phosphorylated chromatin to maintain an adequate balance of kinase and phosphatase activities during mitosis (Murnion et al, 2001, Sugiyama et al, 2002.…”

Section: Chromosome Structure Is Profoundly Altered When Histones Arementioning

confidence: 99%

Histone Hyperacetylation in Mitosis Prevents Sister Chromatid Separation and Produces Chromosome Segregation Defects

Cimini

Mattiuzzo

Torosantucci

et al. 2003

MBoC

167

160

View full text Add to dashboard Cite

Posttranslational modifications of core histones contribute to driving changes in chromatin conformation and compaction. Herein, we investigated the role of histone deacetylation on the mitotic process by inhibiting histone deacetylases shortly before mitosis in human primary fibroblasts. Cells entering mitosis with hyperacetylated histones displayed altered chromatin conformation associated with decreased reactivity to the anti-Ser 10 phospho H3 antibody, increased recruitment of protein phosphatase 1-δ on mitotic chromosomes, and depletion of heterochromatin protein 1 from the centromeric heterochromatin. Inhibition of histone deacetylation before mitosis produced defective chromosome condensation and impaired mitotic progression in living cells, suggesting that improper chromosome condensation may induce mitotic checkpoint activation. In situ hybridization analysis on anaphase cells demonstrated the presence of chromatin bridges, which were caused by persisting cohesion along sister chromatid arms after centromere separation. Thus, the presence of hyperacetylated chromatin during mitosis impairs proper chromosome condensation during the pre-anaphase stages, resulting in poor sister chromatid resolution. Lagging chromosomes consisting of single or paired sisters were also induced by the presence of hyperacetylated histones, indicating that the less constrained centromeric organization associated with heterochromatin protein 1 depletion may promote the attachment of kinetochores to microtubules coming from both poles

show abstract

Aurora-B associated protein phosphatases as negative regulators of kinase activation

Cited by 139 publications

References 42 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

Downregulation of Fer induces PP1 activation and cell-cycle arrest in malignant cells

Histone Hyperacetylation in Mitosis Prevents Sister Chromatid Separation and Produces Chromosome Segregation Defects

Contact Info

Product

Resources

About