Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments

Galina Schevzov, Anthony J. Kee, Bin Wang, Vanessa B. Sequeira, Jeff Hook, Jason D. Coombes, Christine A. Lucas, Justine R. Stehn, Elizabeth A Musgrove, Alexandra Cretu, Richard Assoiane, Thomas Fath, Tamar Hanoch, Rony Seger, Irina Pleines, Benjamin T. Kile, Edna C. Hardeman, Peter W. Gunning

Research output: Contribution to journalArticleResearchpeer-review

Abstract

ERK-regulated cell proliferation requires multiple phosphorylation events catalyzed first by MEK and then by casein kinase 2 (CK2), followed by interaction with importin7 and subsequent nuclear translocation of pERK. We report that genetic manipulation of a core component of the actin filaments of cancer cells, the tropomyosin Tm5NM1, regulates the proliferation of normal cells both in vitro and in vivo. Mouse embryo fibroblasts (MEFs) lacking Tm5NM1, which have reduced proliferative capacity, are insensitive to inhibition of ERK by peptide and small-molecule inhibitors, indicating that ERK is unable to regulate proliferation of these knockout (KO) cells. Treatment of wild-type MEFs with a CK2 inhibitor to block phosphorylation of the nuclear translocation signal in pERK resulted in greatly decreased cell proliferation and a significant reduction in the nuclear translocation of pERK. In contrast, Tm5NM1 KO MEFs, which show reduced nuclear translocation of pERK, were unaffected by inhibition of CK2. This suggested that it is nuclear translocation of CK2-phosphorylated pERK that regulates cell proliferation and this capacity is absent in Tm5NM1 KO cells. Proximity ligation assays confirmed a growth factor-stimulated interaction of pERK with Tm5NM1 and that the interaction of pERK with importin7 is greatly reduced in the Tm5NM1 KO cells.

Original languageEnglish
Pages (from-to)2475-2490
Number of pages16
JournalMolecular Biology of the Cell
Volume26
Issue number13
DOIs
Publication statusPublished - 1 Jul 2015
Externally publishedYes

Cite this

Schevzov, Galina ; Kee, Anthony J. ; Wang, Bin ; Sequeira, Vanessa B. ; Hook, Jeff ; Coombes, Jason D. ; Lucas, Christine A. ; Stehn, Justine R. ; Musgrove, Elizabeth A ; Cretu, Alexandra ; Assoiane, Richard ; Fath, Thomas ; Hanoch, Tamar ; Seger, Rony ; Pleines, Irina ; Kile, Benjamin T. ; Hardeman, Edna C. ; Gunning, Peter W. / Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments. In: Molecular Biology of the Cell. 2015 ; Vol. 26, No. 13. pp. 2475-2490.
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title = "Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments",
abstract = "ERK-regulated cell proliferation requires multiple phosphorylation events catalyzed first by MEK and then by casein kinase 2 (CK2), followed by interaction with importin7 and subsequent nuclear translocation of pERK. We report that genetic manipulation of a core component of the actin filaments of cancer cells, the tropomyosin Tm5NM1, regulates the proliferation of normal cells both in vitro and in vivo. Mouse embryo fibroblasts (MEFs) lacking Tm5NM1, which have reduced proliferative capacity, are insensitive to inhibition of ERK by peptide and small-molecule inhibitors, indicating that ERK is unable to regulate proliferation of these knockout (KO) cells. Treatment of wild-type MEFs with a CK2 inhibitor to block phosphorylation of the nuclear translocation signal in pERK resulted in greatly decreased cell proliferation and a significant reduction in the nuclear translocation of pERK. In contrast, Tm5NM1 KO MEFs, which show reduced nuclear translocation of pERK, were unaffected by inhibition of CK2. This suggested that it is nuclear translocation of CK2-phosphorylated pERK that regulates cell proliferation and this capacity is absent in Tm5NM1 KO cells. Proximity ligation assays confirmed a growth factor-stimulated interaction of pERK with Tm5NM1 and that the interaction of pERK with importin7 is greatly reduced in the Tm5NM1 KO cells.",
author = "Galina Schevzov and Kee, {Anthony J.} and Bin Wang and Sequeira, {Vanessa B.} and Jeff Hook and Coombes, {Jason D.} and Lucas, {Christine A.} and Stehn, {Justine R.} and Musgrove, {Elizabeth A} and Alexandra Cretu and Richard Assoiane and Thomas Fath and Tamar Hanoch and Rony Seger and Irina Pleines and Kile, {Benjamin T.} and Hardeman, {Edna C.} and Gunning, {Peter W.}",
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day = "1",
doi = "10.1091/mbc.E14-10-1453",
language = "English",
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Schevzov, G, Kee, AJ, Wang, B, Sequeira, VB, Hook, J, Coombes, JD, Lucas, CA, Stehn, JR, Musgrove, EA, Cretu, A, Assoiane, R, Fath, T, Hanoch, T, Seger, R, Pleines, I, Kile, BT, Hardeman, EC & Gunning, PW 2015, 'Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments' Molecular Biology of the Cell, vol. 26, no. 13, pp. 2475-2490. https://doi.org/10.1091/mbc.E14-10-1453

Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments. / Schevzov, Galina; Kee, Anthony J.; Wang, Bin; Sequeira, Vanessa B.; Hook, Jeff; Coombes, Jason D.; Lucas, Christine A.; Stehn, Justine R.; Musgrove, Elizabeth A; Cretu, Alexandra ; Assoiane, Richard; Fath, Thomas; Hanoch, Tamar; Seger, Rony; Pleines, Irina ; Kile, Benjamin T.; Hardeman, Edna C.; Gunning, Peter W.

In: Molecular Biology of the Cell, Vol. 26, No. 13, 01.07.2015, p. 2475-2490.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments

AU - Schevzov, Galina

AU - Kee, Anthony J.

AU - Wang, Bin

AU - Sequeira, Vanessa B.

AU - Hook, Jeff

AU - Coombes, Jason D.

AU - Lucas, Christine A.

AU - Stehn, Justine R.

AU - Musgrove, Elizabeth A

AU - Cretu, Alexandra

AU - Assoiane, Richard

AU - Fath, Thomas

AU - Hanoch, Tamar

AU - Seger, Rony

AU - Pleines, Irina

AU - Kile, Benjamin T.

AU - Hardeman, Edna C.

AU - Gunning, Peter W.

PY - 2015/7/1

Y1 - 2015/7/1

N2 - ERK-regulated cell proliferation requires multiple phosphorylation events catalyzed first by MEK and then by casein kinase 2 (CK2), followed by interaction with importin7 and subsequent nuclear translocation of pERK. We report that genetic manipulation of a core component of the actin filaments of cancer cells, the tropomyosin Tm5NM1, regulates the proliferation of normal cells both in vitro and in vivo. Mouse embryo fibroblasts (MEFs) lacking Tm5NM1, which have reduced proliferative capacity, are insensitive to inhibition of ERK by peptide and small-molecule inhibitors, indicating that ERK is unable to regulate proliferation of these knockout (KO) cells. Treatment of wild-type MEFs with a CK2 inhibitor to block phosphorylation of the nuclear translocation signal in pERK resulted in greatly decreased cell proliferation and a significant reduction in the nuclear translocation of pERK. In contrast, Tm5NM1 KO MEFs, which show reduced nuclear translocation of pERK, were unaffected by inhibition of CK2. This suggested that it is nuclear translocation of CK2-phosphorylated pERK that regulates cell proliferation and this capacity is absent in Tm5NM1 KO cells. Proximity ligation assays confirmed a growth factor-stimulated interaction of pERK with Tm5NM1 and that the interaction of pERK with importin7 is greatly reduced in the Tm5NM1 KO cells.

AB - ERK-regulated cell proliferation requires multiple phosphorylation events catalyzed first by MEK and then by casein kinase 2 (CK2), followed by interaction with importin7 and subsequent nuclear translocation of pERK. We report that genetic manipulation of a core component of the actin filaments of cancer cells, the tropomyosin Tm5NM1, regulates the proliferation of normal cells both in vitro and in vivo. Mouse embryo fibroblasts (MEFs) lacking Tm5NM1, which have reduced proliferative capacity, are insensitive to inhibition of ERK by peptide and small-molecule inhibitors, indicating that ERK is unable to regulate proliferation of these knockout (KO) cells. Treatment of wild-type MEFs with a CK2 inhibitor to block phosphorylation of the nuclear translocation signal in pERK resulted in greatly decreased cell proliferation and a significant reduction in the nuclear translocation of pERK. In contrast, Tm5NM1 KO MEFs, which show reduced nuclear translocation of pERK, were unaffected by inhibition of CK2. This suggested that it is nuclear translocation of CK2-phosphorylated pERK that regulates cell proliferation and this capacity is absent in Tm5NM1 KO cells. Proximity ligation assays confirmed a growth factor-stimulated interaction of pERK with Tm5NM1 and that the interaction of pERK with importin7 is greatly reduced in the Tm5NM1 KO cells.

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U2 - 10.1091/mbc.E14-10-1453

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