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 language | English (US) |
|---|---|
| Pages (from-to) | 285-293 |
| Number of pages | 9 |
| Journal | Experimental Cell Research |
| Volume | 296 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jun 10 2004 |
| Externally published | Yes |
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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
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 journal › Article
}
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
C2 - 15149858
AN - SCOPUS:2442570676
VL - 296
SP - 285
EP - 293
JO - Experimental Cell Research
JF - Experimental Cell Research
SN - 0014-4827
IS - 2
ER -