A MEK-independent role for CRAF in mitosis and tumor progression

Ainhoa Mielgo, Laetitia Seguin, Miller Huang, Maria Fernanda Camargo, Sudarshan Anand, Aleksandra Franovic, Sara M. Weis, Sunil J. Advani, Eric A. Murphy, David A. Cheresh

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

RAF kinases regulate cell proliferation and survival and can be dysregulated in tumors 1,2. The role of RAF in cell proliferation has been linked to its ability to activate mitogen-activated protein kinase kinase 1 (MEK) and mitogen-activated protein kinase 1 (ERK). Here we identify a MEK-independent role for RAF in tumor growth. Specifically, in mitotic cells, CRAF becomes phosphorylated on Ser338 and localizes to the mitotic spindle of proliferating tumor cells in vitro as well as in murine tumor models and in biopsies from individuals with cancer. Treatment of tumors with allosteric inhibitors, but not ATP-competitive RAF inhibitors, prevents CRAF phosphorylation on Ser338 and localization to the mitotic spindle and causes cell-cycle arrest at prometaphase. Furthermore, we identify phospho-Ser338 CRAF as a potential biomarker for tumor progression and a surrogate marker for allosteric RAF blockade. Mechanistically, CRAF, but not BRAF, associates with Aurora kinase A (Aurora-A) and Polo-like kinase 1 (Plk1) at the centrosomes and spindle poles during G2/M. Indeed, allosteric or genetic inhibition of phospho-Ser338 CRAF impairs Plk1 activation and accumulation at the kinetochores, causing prometaphase arrest, whereas a phospho-mimetic Ser338D CRAF mutant potentiates Plk1 activation, mitosis and tumor progression in mice. These findings show a previously undefined role for RAF in tumor progression beyond the RAF-MEK-ERK paradigm, opening new avenues for targeting RAF in cancer.

Original languageEnglish (US)
Pages (from-to)1641-1645
Number of pages5
JournalNature Medicine
Volume17
Issue number12
DOIs
StatePublished - Dec 2011
Externally publishedYes

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Mitogen-Activated Protein Kinase Kinases
Mitosis
Tumors
Neoplasms
Prometaphase
Cell proliferation
Spindle Apparatus
Aurora Kinase A
Chemical activation
MAP Kinase Kinase 1
Cells
Phosphorylation
Biopsy
Cell Proliferation
Mitogen-Activated Protein Kinase 1
Spindle Poles
Tumor Biomarkers
Kinetochores
Centrosome
Poles

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Mielgo, A., Seguin, L., Huang, M., Camargo, M. F., Anand, S., Franovic, A., ... Cheresh, D. A. (2011). A MEK-independent role for CRAF in mitosis and tumor progression. Nature Medicine, 17(12), 1641-1645. https://doi.org/10.1038/nm.2464

A MEK-independent role for CRAF in mitosis and tumor progression. / Mielgo, Ainhoa; Seguin, Laetitia; Huang, Miller; Camargo, Maria Fernanda; Anand, Sudarshan; Franovic, Aleksandra; Weis, Sara M.; Advani, Sunil J.; Murphy, Eric A.; Cheresh, David A.

In: Nature Medicine, Vol. 17, No. 12, 12.2011, p. 1641-1645.

Research output: Contribution to journalArticle

Mielgo, A, Seguin, L, Huang, M, Camargo, MF, Anand, S, Franovic, A, Weis, SM, Advani, SJ, Murphy, EA & Cheresh, DA 2011, 'A MEK-independent role for CRAF in mitosis and tumor progression', Nature Medicine, vol. 17, no. 12, pp. 1641-1645. https://doi.org/10.1038/nm.2464
Mielgo A, Seguin L, Huang M, Camargo MF, Anand S, Franovic A et al. A MEK-independent role for CRAF in mitosis and tumor progression. Nature Medicine. 2011 Dec;17(12):1641-1645. https://doi.org/10.1038/nm.2464
Mielgo, Ainhoa ; Seguin, Laetitia ; Huang, Miller ; Camargo, Maria Fernanda ; Anand, Sudarshan ; Franovic, Aleksandra ; Weis, Sara M. ; Advani, Sunil J. ; Murphy, Eric A. ; Cheresh, David A. / A MEK-independent role for CRAF in mitosis and tumor progression. In: Nature Medicine. 2011 ; Vol. 17, No. 12. pp. 1641-1645.
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abstract = "RAF kinases regulate cell proliferation and survival and can be dysregulated in tumors 1,2. The role of RAF in cell proliferation has been linked to its ability to activate mitogen-activated protein kinase kinase 1 (MEK) and mitogen-activated protein kinase 1 (ERK). Here we identify a MEK-independent role for RAF in tumor growth. Specifically, in mitotic cells, CRAF becomes phosphorylated on Ser338 and localizes to the mitotic spindle of proliferating tumor cells in vitro as well as in murine tumor models and in biopsies from individuals with cancer. Treatment of tumors with allosteric inhibitors, but not ATP-competitive RAF inhibitors, prevents CRAF phosphorylation on Ser338 and localization to the mitotic spindle and causes cell-cycle arrest at prometaphase. Furthermore, we identify phospho-Ser338 CRAF as a potential biomarker for tumor progression and a surrogate marker for allosteric RAF blockade. Mechanistically, CRAF, but not BRAF, associates with Aurora kinase A (Aurora-A) and Polo-like kinase 1 (Plk1) at the centrosomes and spindle poles during G2/M. Indeed, allosteric or genetic inhibition of phospho-Ser338 CRAF impairs Plk1 activation and accumulation at the kinetochores, causing prometaphase arrest, whereas a phospho-mimetic Ser338D CRAF mutant potentiates Plk1 activation, mitosis and tumor progression in mice. These findings show a previously undefined role for RAF in tumor progression beyond the RAF-MEK-ERK paradigm, opening new avenues for targeting RAF in cancer.",
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