Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

Cleavage of epidermal growth factor receptor by caspase during apoptosis is independent of its internalization

Oncogene volume 25, pages 1521–1531 (2006)Cite this article

Abstract

Epidermal growth factor receptor (EGFR) plays a critical role in cell proliferation, differentiation, and transformation. EGFR downregulation attenuates its signaling intensity and duration to maintain cellular homeostasis. Here, we report that during apoptosis EGFR is cleaved by activated caspase-3 or related proteases at its C-terminus domain. EGFR downregulation by activation of caspases is neither stimulus- nor cell type-specific. EGFR internalization during apoptosis required dynamin and cholesterol since dominant-negative dynamin (K44A) or cholesterol depletion by methyl-β-cyclodextrin prevented EGFR internalization. However, EGFR downregulation did not require its internalization. The EGFR cleavage fragment was detected in the membrane blebs in addition to the cell pellets. Mutations at the consensus sequence (DXXD) at the C-terminus domain revealed that DVVD1012 and to a lesser extent DNPD1172 may be target sites for active recombinant caspase-3 in vitro and activated caspase-3 or related proteases in vivo. We have detected the N-terminus and C-terminus fragments in vitro and in vivo. A cleavage-deficient EGFR mutant delayed apoptosis process. We conclude that the evolutionarily conserved C-terminus domain of EGFR is the target of caspases and subjected to degradation during apoptosis to shut down its signaling.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9

Similar content being viewed by others

Abbreviations

Ad:

actinomycin D

CA:

camptothecin

CY:

cycloheximide

DEVD (D):

DEVD-fmk,ZD(OMe)-E(OMe)-VD(OMe)-fluoromethylketone

DX:

doxorubicin hydrochloride

EGF:

epidermal growth factor

EGFR:

EGF receptor

EP:

etoposide

FA-fmk (F):

Z-Phe-Ala-fluoromethylketone, a negative control for caspase inhibitor

GM:

GM6001 or Ilomastat, a metalloprotease inhibitor

HRP:

horseradish peroxidase

MβCD (M):

methyl-beta-cyclodextrin

N-EGFR:

anti-EGFR antibody recognizing its N-terminus (EGFR LA22)

Neu:

the second member of the ErB family

PA:

paclitaxel

PDGFR:

platelet-derived growth factor receptor

PU:

puromycin dihydrochloride

rCas-3:

recombinant active human caspase-3

RTKs:

receptor tyrosine kinases

ST:

staurosporine

UVA:

ultraviolet A (315–400 nm)

UVB:

ultraviolet B (280–315 nm)

VAD (V):

VAD-fmk, Z-Val-Ala-Asp (OMe) fluoromethylketone

References

  • Akunda JK, Lao HC, Lee CA, Sessoms AR, Slade RM, Langenbach R . (2004). FASEB J 18: 185–187.

  • Bae SS, Choi JH, Oh YS, Perry DK, Ryu SH, Suh PG . (2001). FEBS Lett 491: 16–20.

  • Benoit V, Chariot A, Delacroix L, Deregowski V, Jacobs N, Merville MP et al. (2004). Cancer Res 64: 2684–2691.

  • Clarke PG . (1990). Anat Embryol (Berl) 181: 195–213.

  • Coleman ML, Sahai EA, Yeo M, Bosch M, Dewar A, Olson MF . (2001). Nat Cell Biol 3: 339–345.

  • Conner SD, Schmid SL . (2003). Nature 422: 37–44.

  • Di Fiore PP, Gill GN . (1999). Curr Opin Cell Biol 11: 483–488.

  • Earnshaw WC, Martins LM, Kaufmann SH . (1999). Annu Rev Biochem 68: 383–424.

  • He YY, Huang JL, Chignell CF . (2004). J Biol Chem 279: 53867–53874.

  • He YY, Huang JL, Gentry JB, Chignell CF . (2003a). J Biol Chem 278: 42457–42465.

  • He YY, Huang JL, Ramirez DC, Chignell CF . (2003b). J Biol Chem 278: 8058–8064.

  • Heldin CH, Westermark B, Wasteson A . (1979). Nature 282: 419–420.

  • Herbst RS, Fukuoka M, Baselga J . (2004). Nat Rev Cancer 4: 956–965.

  • Hunter T . (1997). Cell 88: 333–346.

  • Johnstone RW, Ruefli AA, Lowe SW . (2002). Cell 108: 153–164.

  • Kagaya S, Kitanaka C, Noguchi K, Mochizuki T, Sugiyama A, Asai A et al. (1997). Mol Cell Biol 17: 6736–6745.

  • Kerr JF, Wyllie AH, Currie AR . (1972). Br J Cancer 26: 239–257.

  • Nusbaum P, Laine C, Bouaouina M, Seveau S, Cramer EM, Masse JM et al. (2005). J Biol Chem 280: 5843–5853.

  • Park IC, Park MJ, Choe TB, Jang JJ, Hong SI, Lee SH . (2000). Int J Oncol 16: 1243–1248.

  • Ricci JE, Munoz-Pinedo C, Fitzgerald P, Bailly-Maitre B, Perkins GA, Yadava N et al. (2004). Cell 117: 773–786.

  • Rodal SK, Skretting G, Garred O, Vilhardt F, van Deurs B, Sandvig K . (1999). Mol Biol Cell 10: 961–974.

  • Rowinsky EK . (2004). Annu Rev Med 55: 433–457.

  • Stennicke HR, Renatus M, Meldal M, Salvesen GS . (2000). Biochem J 350(Part 2): 563–568.

  • Subtil A, Gaidarov I, Kobylarz K, Lampson MA, Keen JH, McGraw TE . (1999). Proc Natl Acad Sci USA 96: 6775–6780.

  • Thompson CB . (1995). Science 267: 1456–1462.

  • Tulasne D, Deheuninck J, Lourenco FC, Lamballe F, Ji Z, Leroy C et al. (2004). Mol Cell Biol 24: 10328–10339.

  • Ullrich A, Schlessinger J . (1990). Cell 61: 203–212.

  • Wyllie AH, Kerr JF, Currie AR . (1980). Int Rev Cytol 68: 251–306.

  • Zhuang S, Ouedraogo GD, Kochevar IE . (2003). Oncogene 22: 4413–4424.

Download references

Acknowledgements

The research was supported by the intramural research program of the NIH, the NIEHS. We thank Jeff Reece for his help with confocal microscopy measurements, and Carl Bortner for his help with flow cytometry. We thank John O'Bryan for helpful discussion. We are also grateful to Drs Ann Motten and Jie Liu for critical reading of the manuscript.

Author information

Authors and Affiliations

  1. Laboratory of Pharmacology and Chemistry, NIEHS, National Institute of Health, Research Triangle Park, North Carolina, USA

    Y-Y He, J-L Huang & C F Chignell

Authors
  1. Y-Y He
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J-L Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C F Chignell
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Y-Y He.

Rights and permissions

Reprints and permissions

About this article

Cite this article

He, YY., Huang, JL. & Chignell, C. Cleavage of epidermal growth factor receptor by caspase during apoptosis is independent of its internalization. Oncogene 25, 1521–1531 (2006). https://doi.org/10.1038/sj.onc.1209184

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1209184

Keywords

This article is cited by

Search

Advanced search

Quick links