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Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts.

Publication ,  Journal Article
Smith, JJ; Evans, EK; Murakami, M; Moyer, MB; Moseley, MA; Vande Woude, G; Kornbluth, S
Published in: J Cell Biol
December 25, 2000

Many of the biochemical reactions of apoptotic cell death, including mitochondrial cytochrome c release and caspase activation, can be reconstituted in cell-free extracts derived from Xenopus eggs. In addition, because caspase activation does not occur until the egg extract has been incubated for several hours on the bench, upstream signaling processes occurring before full apoptosis are rendered accessible to biochemical manipulation. We reported previously that the adaptor protein Crk is required for apoptotic signaling in egg extracts (Evans, E.K., W. Lu, S.L. Strum, B.J. Mayer, and S. Kornbluth. 1997. EMBO (Eur. Mol. Biol. Organ.) J. 16:230-241). Moreover, we demonstrated that removal of Crk Src homology (SH)2 or SH3 interactors from the extracts prevented apoptosis. We now report the finding that the relevant Crk SH2-interacting protein, important for apoptotic signaling in the extract, is the well-known cell cycle regulator, Wee1. We have demonstrated a specific interaction between tyrosine-phosphorylated Wee1 and the Crk SH2 domain and have shown that recombinant Wee1 can restore apoptosis to an extract depleted of SH2 interactors. Moreover, exogenous Wee1 accelerated apoptosis in egg extracts, and this acceleration was largely dependent on the presence of endogenous Crk protein. As other Cdk inhibitors, such as roscovitine and Myt1, did not act like Wee1 to accelerate apoptosis, we propose that Wee1-Crk complexes signal in a novel apoptotic pathway, which may be unrelated to Wee1's role as a cell cycle regulator.

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Published In

J Cell Biol

DOI

ISSN

0021-9525

Publication Date

December 25, 2000

Volume

151

Issue

7

Start / End Page

1391 / 1400

Location

United States

Related Subject Headings

  • src Homology Domains
  • Xenopus laevis
  • Xenopus Proteins
  • Signal Transduction
  • Roscovitine
  • Purines
  • Proto-Oncogene Proteins c-crk
  • Proto-Oncogene Proteins
  • Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases
 

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Smith, J. J., Evans, E. K., Murakami, M., Moyer, M. B., Moseley, M. A., Vande Woude, G., & Kornbluth, S. (2000). Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts. J Cell Biol, 151(7), 1391–1400. https://doi.org/10.1083/jcb.151.7.1391
Smith, J. J., E. K. Evans, M. Murakami, M. B. Moyer, M. A. Moseley, G. Vande Woude, and S. Kornbluth. “Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts.J Cell Biol 151, no. 7 (December 25, 2000): 1391–1400. https://doi.org/10.1083/jcb.151.7.1391.
Smith JJ, Evans EK, Murakami M, Moyer MB, Moseley MA, Vande Woude G, et al. Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts. J Cell Biol. 2000 Dec 25;151(7):1391–400.
Smith, J. J., et al. “Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts.J Cell Biol, vol. 151, no. 7, Dec. 2000, pp. 1391–400. Pubmed, doi:10.1083/jcb.151.7.1391.
Smith JJ, Evans EK, Murakami M, Moyer MB, Moseley MA, Vande Woude G, Kornbluth S. Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts. J Cell Biol. 2000 Dec 25;151(7):1391–1400.

Published In

J Cell Biol

DOI

ISSN

0021-9525

Publication Date

December 25, 2000

Volume

151

Issue

7

Start / End Page

1391 / 1400

Location

United States

Related Subject Headings

  • src Homology Domains
  • Xenopus laevis
  • Xenopus Proteins
  • Signal Transduction
  • Roscovitine
  • Purines
  • Proto-Oncogene Proteins c-crk
  • Proto-Oncogene Proteins
  • Protein-Tyrosine Kinases
  • Protein Serine-Threonine Kinases