Abstract
A hybrid receptor molecule composed of the extracellular ligand-binding domain of the human insulin receptor and the transmembrane and cytoplasmic (protein-tyrosine kinase) domains of the chicken sarcoma virus UR2 transforming protein p68gag-ros has been constructed and expressed in Chinese hamster ovary (CHO) cells. The hybrid is processed normally into alpha and hybrid beta subunits, is expressed on the cell surface at high levels, and binds insulin with near-wild-type affinity. Furthermore, insulin stimulates the phosphorylation on tyrosine residues of the hybrid beta subunit in vivo and the phosphorylation of an exogenous substrate [poly(Glu,Tyr)] in vitro. Thus the hybrid is capable of heterologous transmembrane signaling. However, the hybrid mediates neither the insulin-activated uptake of 2-deoxyglucose nor the incorporation of [3H]thymidine into DNA, suggesting that the physiological response(s) mediated by ligand-activated protein-tyrosine kinases may utilize distinct intracellular mechanisms for postreceptor signaling.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Balduzzi P. C., Notter M. F., Morgan H. R., Shibuya M. Some biological properties of two new avian sarcoma viruses. J Virol. 1981 Oct;40(1):268–275. doi: 10.1128/jvi.40.1.268-275.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cooper J. A., Hunter T. Changes in protein phosphorylation in Rous sarcoma virus-transformed chicken embryo cells. Mol Cell Biol. 1981 Feb;1(2):165–178. doi: 10.1128/mcb.1.2.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ebina Y., Ellis L., Jarnagin K., Edery M., Graf L., Clauser E., Ou J. H., Masiarz F., Kan Y. W., Goldfine I. D. The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling. Cell. 1985 Apr;40(4):747–758. doi: 10.1016/0092-8674(85)90334-4. [DOI] [PubMed] [Google Scholar]
- Ellis L., Clauser E., Morgan D. O., Edery M., Roth R. A., Rutter W. J. Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose. Cell. 1986 Jun 6;45(5):721–732. doi: 10.1016/0092-8674(86)90786-5. [DOI] [PubMed] [Google Scholar]
- Ellis L., Morgan D. O., Koshland D. E., Jr, Clauser E., Moe G. R., Bollag G., Roth R. A., Rutter W. J. Linking functional domains of the human insulin receptor with the bacterial aspartate receptor. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8137–8141. doi: 10.1073/pnas.83.21.8137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feldman R. A., Wang L. H., Hanafusa H., Balduzzi P. C. Avian sarcoma virus UR2 encodes a transforming protein which is associated with a unique protein kinase activity. J Virol. 1982 Apr;42(1):228–236. doi: 10.1128/jvi.42.1.228-236.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
- Kahn C. R. The molecular mechanism of insulin action. Annu Rev Med. 1985;36:429–451. doi: 10.1146/annurev.me.36.020185.002241. [DOI] [PubMed] [Google Scholar]
- Kasuga M., Zick Y., Blith D. L., Karlsson F. A., Häring H. U., Kahn C. R. Insulin stimulation of phosphorylation of the beta subunit of the insulin receptor. Formation of both phosphoserine and phosphotyrosine. J Biol Chem. 1982 Sep 10;257(17):9891–9894. [PubMed] [Google Scholar]
- Morgan D. O., Ho L., Korn L. J., Roth R. A. Insulin action is blocked by a monoclonal antibody that inhibits the insulin receptor kinase. Proc Natl Acad Sci U S A. 1986 Jan;83(2):328–332. doi: 10.1073/pnas.83.2.328. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morgan D. O., Roth R. A. Acute insulin action requires insulin receptor kinase activity: introduction of an inhibitory monoclonal antibody into mammalian cells blocks the rapid effects of insulin. Proc Natl Acad Sci U S A. 1987 Jan;84(1):41–45. doi: 10.1073/pnas.84.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morgan D. O., Roth R. A. Mapping surface structures of the human insulin receptor with monoclonal antibodies: localization of main immunogenic regions to the receptor kinase domain. Biochemistry. 1986 Mar 25;25(6):1364–1371. doi: 10.1021/bi00354a026. [DOI] [PubMed] [Google Scholar]
- Neckameyer W. S., Shibuya M., Hsu M. T., Wang L. H. Proto-oncogene c-ros codes for a molecule with structural features common to those of growth factor receptors and displays tissue specific and developmentally regulated expression. Mol Cell Biol. 1986 May;6(5):1478–1486. doi: 10.1128/mcb.6.5.1478. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Neckameyer W. S., Wang L. H. Nucleotide sequence of avian sarcoma virus UR2 and comparison of its transforming gene with other members of the tyrosine protein kinase oncogene family. J Virol. 1985 Mar;53(3):879–884. doi: 10.1128/jvi.53.3.879-884.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Podskalny J., McElduff A., Gorden P. Insulin receptors on Chinese hamster ovary (CHO) cells: altered insulin binding to glycosylation mutants. Biochem Biophys Res Commun. 1984 Nov 30;125(1):70–75. doi: 10.1016/s0006-291x(84)80335-6. [DOI] [PubMed] [Google Scholar]
- Roth R. A., Cassell D. J., Wong K. Y., Maddux B. A., Goldfine I. D. Monoclonal antibodies to the human insulin receptor block insulin binding and inhibit insulin action. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7312–7316. doi: 10.1073/pnas.79.23.7312. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shibuya M., Hanafusa H., Balduzzi P. C. Cellular sequences related to three new onc genes of avian sarcoma virus (fps, yes, and ros) and their expression in normal and transformed cells. J Virol. 1982 Apr;42(1):143–152. doi: 10.1128/jvi.42.1.143-152.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
- Ullrich A., Bell J. R., Chen E. Y., Herrera R., Petruzzelli L. M., Dull T. J., Gray A., Coussens L., Liao Y. C., Tsubokawa M. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes. 1985 Feb 28-Mar 6Nature. 313(6005):756–761. doi: 10.1038/313756a0. [DOI] [PubMed] [Google Scholar]
- Wang L. H., Hanafusa H., Notter M. F., Balduzzi P. C. Genetic structure and transforming sequence of avian sarcoma virus UR2. J Virol. 1982 Mar;41(3):833–841. doi: 10.1128/jvi.41.3.833-841.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]