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.
N1 - Funding Information:
We gratefully acknowledge Dr. Stephen Hughes (NCI) for providing the RCAN retroviral vectors used in these studies. We also thank Shari Meeker, Laura Sikkink, Annabeth Fieck, and Emily Ward for their technical contributions, and Drs. Ze Wang, Jill Reiter, Mike McManus, and Julie Boerner for their technical assistance and helpful discussions. This work was supported by the National Institutes of Health (CA79808).
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
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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
SN - 0014-4827
VL - 296
SP - 285
EP - 293
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -