Wnt-5a-induced phosphorylation of DARPP-32 inhibits breast cancer cell migration in a CREB-dependent manner.
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1470124
- author
- Hansen, Christian LU ; Howlin, Jillian LU ; Tengholm, Anders ; Dyachok, Oleg ; Vogel, Wolfgang F ; Nairn, Angus C ; Greengard, Paul and Andersson, Tommy LU
- organization
- publishing date
- 2009
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 284
- pages
- 27533 - 27543
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- wos:000270232300055
- pmid:19651774
- scopus:70350439474
- pmid:19651774
- 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
- 2016-04-04 08:11:48
- date last changed
- 2022-04-07 23:40:21
@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 = {{American Society for Biochemistry and Molecular Biology}},
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}},
doi = {{10.1074/jbc.M109.048884}},
volume = {{284}},
year = {{2009}},
}