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Wnt-5a-induced phosphorylation of DARPP-32 inhibits breast cancer cell migration in a CREB-dependent manner.

Hansen, Christian LU ; Howlin, Jillian LU ; Tengholm, Anders; Dyachok, Oleg; Vogel, Wolfgang F; Nairn, Angus C; Greengard, Paul and Andersson, Tommy LU (2009) In Journal of Biological Chemistry 284. p.27533-27543
Abstract
Tumour cell migration plays a central role in the process of cancer metastasis. We recently identified dopamine and cAMP regulated phospho-protein of 32 kDa (DARPP-32) as an anti-migratory phospho-protein in breast cancer cells. Here we link this effect of DARPP-32 to Wnt-5a signalling by demonstrating that recombinant Wnt-5a triggers cAMP elevation at the plasma membrane and Thr34-DARPP-32 phosphorylation in MCF-7 cells. In agreement, both protein kinase A (PKA) inhibitors and siRNA-mediated knock down of Frizzled-3 receptor or Galphas expression abolished Wnt-5a-induced phosphorylation of DARPP-32. Furthermore, Wnt-5a induced DARPP-32 dependent inhibition of MCF-7 cell migration. Phospho-Thr34-DARPP-32 interacted with protein... (More)
Tumour cell migration plays a central role in the process of cancer metastasis. We recently identified dopamine and cAMP regulated phospho-protein of 32 kDa (DARPP-32) as an anti-migratory phospho-protein in breast cancer cells. Here we link this effect of DARPP-32 to Wnt-5a signalling by demonstrating that recombinant Wnt-5a triggers cAMP elevation at the plasma membrane and Thr34-DARPP-32 phosphorylation in MCF-7 cells. In agreement, both protein kinase A (PKA) inhibitors and siRNA-mediated knock down of Frizzled-3 receptor or Galphas expression abolished Wnt-5a-induced phosphorylation of DARPP-32. Furthermore, Wnt-5a induced DARPP-32 dependent inhibition of MCF-7 cell migration. Phospho-Thr34-DARPP-32 interacted with protein phosphatase-1 (PP1) and potentiated the Wnt-5a-mediated phosphorylation of CREB, a well-known PP1 substrate, but had no effect on CREB phosphorylation by itself. Moreover, inhibition of the Wnt-5a/DARPP-32/CREB pathway, by expression of dominant negative CREB (DN-CREB), diminished the anti-migratory effect of Wnt-5a-induced phospho-Thr34-DARPP-32. Phalloidin-staining revealed that that the presence of phospho-Thr34-DARPP-32 in MCF-7 cells results in reduced filopodia formation. In accordance, the activity of the Rho GTPase Cdc42, known to be crucial for filopodia formation, was reduced in MCF-7 cells expressing phospho-Thr34-DARPP-32. The effects of DARPP-32 on cell migration and filopodia formation could be reversed in T47D breast cancer cells that were depleted of their endogenous DARPP-32 by siRNA targeting. Consequently, Wnt-5a activates a Frizzled-3/Galphas /cAMP/PKA signalling pathway that triggers a DARPP-32- and CREB-dependent anti-migratory response in breast cancer cells, representing a novel mechanism whereby Wnt-5a can inhibit breast cancer cell migration. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
284
pages
27533 - 27543
publisher
ASBMB
external identifiers
  • wos:000270232300055
  • pmid:19651774
  • scopus:70350439474
ISSN
1083-351X
DOI
10.1074/jbc.M109.048884
language
English
LU publication?
yes
id
9c7c6051-0e3a-4e51-8168-4c4b6216c63c (old id 1470124)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/19651774?dopt=Abstract
date added to LUP
2009-09-04 11:13:25
date last changed
2017-06-04 04:32:32
@article{9c7c6051-0e3a-4e51-8168-4c4b6216c63c,
  abstract     = {Tumour cell migration plays a central role in the process of cancer metastasis. We recently identified dopamine and cAMP regulated phospho-protein of 32 kDa (DARPP-32) as an anti-migratory phospho-protein in breast cancer cells. Here we link this effect of DARPP-32 to Wnt-5a signalling by demonstrating that recombinant Wnt-5a triggers cAMP elevation at the plasma membrane and Thr34-DARPP-32 phosphorylation in MCF-7 cells. In agreement, both protein kinase A (PKA) inhibitors and siRNA-mediated knock down of Frizzled-3 receptor or Galphas expression abolished Wnt-5a-induced phosphorylation of DARPP-32. Furthermore, Wnt-5a induced DARPP-32 dependent inhibition of MCF-7 cell migration. Phospho-Thr34-DARPP-32 interacted with protein phosphatase-1 (PP1) and potentiated the Wnt-5a-mediated phosphorylation of CREB, a well-known PP1 substrate, but had no effect on CREB phosphorylation by itself. Moreover, inhibition of the Wnt-5a/DARPP-32/CREB pathway, by expression of dominant negative CREB (DN-CREB), diminished the anti-migratory effect of Wnt-5a-induced phospho-Thr34-DARPP-32. Phalloidin-staining revealed that that the presence of phospho-Thr34-DARPP-32 in MCF-7 cells results in reduced filopodia formation. In accordance, the activity of the Rho GTPase Cdc42, known to be crucial for filopodia formation, was reduced in MCF-7 cells expressing phospho-Thr34-DARPP-32. The effects of DARPP-32 on cell migration and filopodia formation could be reversed in T47D breast cancer cells that were depleted of their endogenous DARPP-32 by siRNA targeting. Consequently, Wnt-5a activates a Frizzled-3/Galphas /cAMP/PKA signalling pathway that triggers a DARPP-32- and CREB-dependent anti-migratory response in breast cancer cells, representing a novel mechanism whereby Wnt-5a can inhibit breast cancer cell migration.},
  author       = {Hansen, Christian and Howlin, Jillian and Tengholm, Anders and Dyachok, Oleg and Vogel, Wolfgang F and Nairn, Angus C and Greengard, Paul and Andersson, Tommy},
  issn         = {1083-351X},
  language     = {eng},
  pages        = {27533--27543},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Wnt-5a-induced phosphorylation of DARPP-32 inhibits breast cancer cell migration in a CREB-dependent manner.},
  url          = {http://dx.doi.org/10.1074/jbc.M109.048884},
  volume       = {284},
  year         = {2009},
}