Tyrosyl phosphorylation of KRAS stalls GTPase cycle via alteration of switch I and II conformation

Yoshihito Kano, Teklab Gebregiworgis, Christopher B. Marshall, Nikolina Radulovich, Betty P.K. Poon, Jonathan St-Germain, Jonathan D. Cook, Ivette Valencia-Sama, Benjamin M.M. Grant, Silvia Gabriela Herrera, Jinmin Miao, Brian Raught, Meredith S. Irwin, Jeffrey E. Lee, Jen Jen Yeh, Zhong Yin Zhang, Ming Sound Tsao, Mitsuhiko Ikura, Michael Ohh

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6 Scopus citations


Deregulation of the RAS GTPase cycle due to mutations in the three RAS genes is commonly associated with cancer development. Protein tyrosine phosphatase SHP2 promotes RAF-to-MAPK signaling pathway and is an essential factor in RAS-driven oncogenesis. Despite the emergence of SHP2 inhibitors for the treatment of cancers harbouring mutant KRAS, the mechanism underlying SHP2 activation of KRAS signaling remains unclear. Here we report tyrosyl-phosphorylation of endogenous RAS and demonstrate that KRAS phosphorylation via Src on Tyr32 and Tyr64 alters the conformation of switch I and II regions, which stalls multiple steps of the GTPase cycle and impairs binding to effectors. In contrast, SHP2 dephosphorylates KRAS, a process that is required to maintain dynamic canonical KRAS GTPase cycle. Notably, Src- and SHP2-mediated regulation of KRAS activity extends to oncogenic KRAS and the inhibition of SHP2 disrupts the phosphorylation cycle, shifting the equilibrium of the GTPase cycle towards the stalled ‘dark state’.

Original languageEnglish (US)
Article number224
JournalNature communications
Issue number1
StatePublished - Dec 1 2019


ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Kano, Y., Gebregiworgis, T., Marshall, C. B., Radulovich, N., Poon, B. P. K., St-Germain, J., Cook, J. D., Valencia-Sama, I., Grant, B. M. M., Herrera, S. G., Miao, J., Raught, B., Irwin, M. S., Lee, J. E., Yeh, J. J., Zhang, Z. Y., Tsao, M. S., Ikura, M., & Ohh, M. (2019). Tyrosyl phosphorylation of KRAS stalls GTPase cycle via alteration of switch I and II conformation. Nature communications, 10(1), [224]. https://doi.org/10.1038/s41467-018-08115-8