Membrane localization of v-ErbB is required but not sufficient for ligand-independent transformation

Andrew J. Danielsen, Trace A. Christensen, Courtney A. Lovejoy, Margaret A. Adelsman, Denise C. Connolly, Nita J. Maihle

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The v-ErbB retroviral oncogene is a transduced, mutated copy of the avian EGF receptor gene, and its expression is sufficient to induce tumor formation in vivo. The structural alterations that release the oncogenic potential of the v-ErbB oncogene are similar to EGFR gene mutations described in human tumors. Thus, the study of v-ErbB tumor biology offers a useful model through which we can gain insight into the mechanism of EGFR-induced malignancies. Despite years of study, however, questions remain regarding the domains of v-ErbB required for oncogenicity. We sought to clarify the role of the transmembrane domain of v-ErbB during transformation using S3-v-ErbB, an acutely transforming retroviral oncogene isolated from avian sarcomas. Infection of primary fibroblasts with a retroviral vector containing S3-v-ErbB results in the formation of a transformation-associated phosphoprotein signaling complex, soft agar colony formation, and the rapid induction of highly vascularized sarcomas in vivo. To address contribution of the transmembrane domain of S3-v-ErbB during these processes, we constructed a mutant version of this oncogene with a precise deletion in this domain. Specifically, the S3-v-ErbB-TM- mutant was created through an in-frame deletion of the entire transmembrane domain. Primary fibroblasts expressing this S3-v-ErbB-TM- mutant fail to form a characteristic transformation-associated phosphoprotein complex and do not grow in an anchorage-independent manner. In addition, day-old chicks injected with a helper-independent retrovirus expressing the S3-v-ErbB-TM- mutant exhibit only limited tumor formation in vivo. These results demonstrate that the transmembrane domain and, consequently membrane localization, are essential for S3-v-ErbB-mediated transformation.

Original languageEnglish (US)
Pages (from-to)285-293
Number of pages9
JournalExperimental Cell Research
Volume296
Issue number2
DOIs
StatePublished - Jun 10 2004
Externally publishedYes

Fingerprint

erbB-1 Genes
Ligands
Membranes
Phosphoproteins
Neoplasms
Oncogenes
Avian Sarcoma
Fibroblasts
Retroviridae
Epidermal Growth Factor Receptor
Sarcoma
Agar
Gene Expression
Mutation
Infection

Keywords

  • AEV
  • Avian erythroblastosis virus
  • CEF
  • Chick embryo fibroblasts
  • EGF/ErbB receptors
  • EGFR
  • Epidermal growth factor receptor
  • Nuclear localization
  • Transmembrane domain
  • v-ErbB

ASJC Scopus subject areas

  • Cell Biology

Cite this

Danielsen, A. J., Christensen, T. A., Lovejoy, C. A., Adelsman, M. A., Connolly, D. C., & Maihle, N. J. (2004). Membrane localization of v-ErbB is required but not sufficient for ligand-independent transformation. Experimental Cell Research, 296(2), 285-293. https://doi.org/10.1016/j.yexcr.2004.01.023

Membrane localization of v-ErbB is required but not sufficient for ligand-independent transformation. / Danielsen, Andrew J.; Christensen, Trace A.; Lovejoy, Courtney A.; Adelsman, Margaret A.; Connolly, Denise C.; Maihle, Nita J.

In: Experimental Cell Research, Vol. 296, No. 2, 10.06.2004, p. 285-293.

Research output: Contribution to journalArticle

Danielsen, AJ, Christensen, TA, Lovejoy, CA, Adelsman, MA, Connolly, DC & Maihle, NJ 2004, 'Membrane localization of v-ErbB is required but not sufficient for ligand-independent transformation', Experimental Cell Research, vol. 296, no. 2, pp. 285-293. https://doi.org/10.1016/j.yexcr.2004.01.023
Danielsen, Andrew J. ; Christensen, Trace A. ; Lovejoy, Courtney A. ; Adelsman, Margaret A. ; Connolly, Denise C. ; Maihle, Nita J. / Membrane localization of v-ErbB is required but not sufficient for ligand-independent transformation. In: Experimental Cell Research. 2004 ; Vol. 296, No. 2. pp. 285-293.
@article{6f7d1a4035a64f91884e7e61359d91c7,
title = "Membrane localization of v-ErbB is required but not sufficient for ligand-independent transformation",
abstract = "The v-ErbB retroviral oncogene is a transduced, mutated copy of the avian EGF receptor gene, and its expression is sufficient to induce tumor formation in vivo. The structural alterations that release the oncogenic potential of the v-ErbB oncogene are similar to EGFR gene mutations described in human tumors. Thus, the study of v-ErbB tumor biology offers a useful model through which we can gain insight into the mechanism of EGFR-induced malignancies. Despite years of study, however, questions remain regarding the domains of v-ErbB required for oncogenicity. We sought to clarify the role of the transmembrane domain of v-ErbB during transformation using S3-v-ErbB, an acutely transforming retroviral oncogene isolated from avian sarcomas. Infection of primary fibroblasts with a retroviral vector containing S3-v-ErbB results in the formation of a transformation-associated phosphoprotein signaling complex, soft agar colony formation, and the rapid induction of highly vascularized sarcomas in vivo. To address contribution of the transmembrane domain of S3-v-ErbB during these processes, we constructed a mutant version of this oncogene with a precise deletion in this domain. Specifically, the S3-v-ErbB-TM- mutant was created through an in-frame deletion of the entire transmembrane domain. Primary fibroblasts expressing this S3-v-ErbB-TM- mutant fail to form a characteristic transformation-associated phosphoprotein complex and do not grow in an anchorage-independent manner. In addition, day-old chicks injected with a helper-independent retrovirus expressing the S3-v-ErbB-TM- mutant exhibit only limited tumor formation in vivo. These results demonstrate that the transmembrane domain and, consequently membrane localization, are essential for S3-v-ErbB-mediated transformation.",
keywords = "AEV, Avian erythroblastosis virus, CEF, Chick embryo fibroblasts, EGF/ErbB receptors, EGFR, Epidermal growth factor receptor, Nuclear localization, Transmembrane domain, v-ErbB",
author = "Danielsen, {Andrew J.} and Christensen, {Trace A.} and Lovejoy, {Courtney A.} and Adelsman, {Margaret A.} and Connolly, {Denise C.} and Maihle, {Nita J.}",
year = "2004",
month = "6",
day = "10",
doi = "10.1016/j.yexcr.2004.01.023",
language = "English (US)",
volume = "296",
pages = "285--293",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Membrane localization of v-ErbB is required but not sufficient for ligand-independent transformation

AU - Danielsen, Andrew J.

AU - Christensen, Trace A.

AU - Lovejoy, Courtney A.

AU - Adelsman, Margaret A.

AU - Connolly, Denise C.

AU - Maihle, Nita J.

PY - 2004/6/10

Y1 - 2004/6/10

N2 - The v-ErbB retroviral oncogene is a transduced, mutated copy of the avian EGF receptor gene, and its expression is sufficient to induce tumor formation in vivo. The structural alterations that release the oncogenic potential of the v-ErbB oncogene are similar to EGFR gene mutations described in human tumors. Thus, the study of v-ErbB tumor biology offers a useful model through which we can gain insight into the mechanism of EGFR-induced malignancies. Despite years of study, however, questions remain regarding the domains of v-ErbB required for oncogenicity. We sought to clarify the role of the transmembrane domain of v-ErbB during transformation using S3-v-ErbB, an acutely transforming retroviral oncogene isolated from avian sarcomas. Infection of primary fibroblasts with a retroviral vector containing S3-v-ErbB results in the formation of a transformation-associated phosphoprotein signaling complex, soft agar colony formation, and the rapid induction of highly vascularized sarcomas in vivo. To address contribution of the transmembrane domain of S3-v-ErbB during these processes, we constructed a mutant version of this oncogene with a precise deletion in this domain. Specifically, the S3-v-ErbB-TM- mutant was created through an in-frame deletion of the entire transmembrane domain. Primary fibroblasts expressing this S3-v-ErbB-TM- mutant fail to form a characteristic transformation-associated phosphoprotein complex and do not grow in an anchorage-independent manner. In addition, day-old chicks injected with a helper-independent retrovirus expressing the S3-v-ErbB-TM- mutant exhibit only limited tumor formation in vivo. These results demonstrate that the transmembrane domain and, consequently membrane localization, are essential for S3-v-ErbB-mediated transformation.

AB - The v-ErbB retroviral oncogene is a transduced, mutated copy of the avian EGF receptor gene, and its expression is sufficient to induce tumor formation in vivo. The structural alterations that release the oncogenic potential of the v-ErbB oncogene are similar to EGFR gene mutations described in human tumors. Thus, the study of v-ErbB tumor biology offers a useful model through which we can gain insight into the mechanism of EGFR-induced malignancies. Despite years of study, however, questions remain regarding the domains of v-ErbB required for oncogenicity. We sought to clarify the role of the transmembrane domain of v-ErbB during transformation using S3-v-ErbB, an acutely transforming retroviral oncogene isolated from avian sarcomas. Infection of primary fibroblasts with a retroviral vector containing S3-v-ErbB results in the formation of a transformation-associated phosphoprotein signaling complex, soft agar colony formation, and the rapid induction of highly vascularized sarcomas in vivo. To address contribution of the transmembrane domain of S3-v-ErbB during these processes, we constructed a mutant version of this oncogene with a precise deletion in this domain. Specifically, the S3-v-ErbB-TM- mutant was created through an in-frame deletion of the entire transmembrane domain. Primary fibroblasts expressing this S3-v-ErbB-TM- mutant fail to form a characteristic transformation-associated phosphoprotein complex and do not grow in an anchorage-independent manner. In addition, day-old chicks injected with a helper-independent retrovirus expressing the S3-v-ErbB-TM- mutant exhibit only limited tumor formation in vivo. These results demonstrate that the transmembrane domain and, consequently membrane localization, are essential for S3-v-ErbB-mediated transformation.

KW - AEV

KW - Avian erythroblastosis virus

KW - CEF

KW - Chick embryo fibroblasts

KW - EGF/ErbB receptors

KW - EGFR

KW - Epidermal growth factor receptor

KW - Nuclear localization

KW - Transmembrane domain

KW - v-ErbB

UR - http://www.scopus.com/inward/record.url?scp=2442570676&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2442570676&partnerID=8YFLogxK

U2 - 10.1016/j.yexcr.2004.01.023

DO - 10.1016/j.yexcr.2004.01.023

M3 - Article

VL - 296

SP - 285

EP - 293

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 2

ER -