Abstract
Regulation of the cytoplasmic protein changes during myeloid cell differentiation has been analyzed with two-dimensional gel electrophoresis and differentiation-defective cell mutants. The cells studied include a clone of myeloid leukemia cells (clone 11) that can be induced to differentiate to macrophages by the protein inducer MGI and the steroid dexamethasone (Dex) and mutant clones that were inducible for differentiation to macrophages by MGI but not by Dex. The mutants were not defective in the specific binding of [3H]Dex to cytoplasmic receptors or in the transport and nuclear binding of the receptor--steroid complex. The protein patterns in the mutants showed both specific constitutive protein changes and nonresponding proteins. Twenty-one percent of the Dex-induced protein changes and 2% of the MGI-induced protein changes in clone 11 were constitutively expressed in the mutants. In addition, 28% of the proteins that responded to Dex in clone 11 did not respond to Dex in the mutants, whereas only 4% of the proteins that responded to MGI in clone 11 did not respond to MGI. The higher percentage of constitutive changes was thus associated with a larger defect in induction. The proteins with an abnormal response to Dex still showed a normal response to MGI, and the constitutive changes and nonresponding proteins were different for the two inducers. It is suggested that specific constitutive protein changes expressed by the mutants produced an asynchrony in the developmental program, resulting in a defective response to Dex and to MGI, and that this may apply to other inducers and developmental programs.
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- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- Cox R. P., Elson N. A., Tu S. H., Griffin M. J. Hormonal induction of alkaline phosphatase activity by an increase in catalytic efficiency of the enzyme. J Mol Biol. 1971 May 28;58(1):197–215. doi: 10.1016/0022-2836(71)90241-5. [DOI] [PubMed] [Google Scholar]
- Friend C. The phenomenon of differentiation in murine erythroleukemic cells. Harvey Lect. 1978;72:253–281. [PubMed] [Google Scholar]
- Hoffman-Liebermann B., Sachs L. Regulation of actin and other proteins in the differentiation of myeloid leukemic cells. Cell. 1978 Aug;14(4):825–834. doi: 10.1016/0092-8674(78)90338-0. [DOI] [PubMed] [Google Scholar]
- Ivarie R. D., O'Farrell P. H. The glucocorticoid domain: steroid-mediated changes in the rate of synthesis of rat hepatoma proteins. Cell. 1978 Jan;13(1):41–55. doi: 10.1016/0092-8674(78)90136-8. [DOI] [PubMed] [Google Scholar]
- Krystosek A., Sachs L. Steroid hormone receptors and the differentiation of myeloid leukemic cells. J Cell Physiol. 1977 Sep;92(3):345–352. doi: 10.1002/jcp.1040920303. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
- Liebermann D., Hoffman-Liebermann B., Sachs L. Molecular dissection of differentiation in normal and leukemic myeloblasts: separately programmed pathways of gene expression. Dev Biol. 1980 Sep;79(1):46–63. doi: 10.1016/0012-1606(80)90072-x. [DOI] [PubMed] [Google Scholar]
- Linser P., Moscona A. A. Induction of glutamine synthetase in embryonic neural retina: localization in Müller fibers and dependence on cell interactions. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6476–6480. doi: 10.1073/pnas.76.12.6476. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lotem J., Sachs L. Different blocks in the differentiation of myeloid leukemic cells. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3507–3511. doi: 10.1073/pnas.71.9.3507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lotem J., Sachs L. Genetic dissection of the control of normal differentiation in myeloid leukemic cells. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5554–5558. doi: 10.1073/pnas.74.12.5554. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lotem J., Sachs L. In vivo induction of normal differentiation in myeloid leukemia cells. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3781–3785. doi: 10.1073/pnas.75.8.3781. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lotem J., Sachs L. Induction of specific changes in the surface membrane of myeloid leukemic cells by steroid hormones. Int J Cancer. 1975 May 15;15(5):731–740. doi: 10.1002/ijc.2910150504. [DOI] [PubMed] [Google Scholar]
- Marks P. A., Rifkind R. A. Erythroleukemic differentiation. Annu Rev Biochem. 1978;47:419–448. doi: 10.1146/annurev.bi.47.070178.002223. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
- Sachs L. Constitutive uncoupling of pathways of gene expression that control growth and differentiation in myeloid leukemia: a model for the origin and progression of malignancy. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6152–6156. doi: 10.1073/pnas.77.10.6152. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sachs L. Control of normal cell differentiation and the phenotypic reversion of malignancy in myeloid leukaemia. Nature. 1978 Aug 10;274(5671):535–539. doi: 10.1038/274535a0. [DOI] [PubMed] [Google Scholar]
- Sela B. A., Sachs L. Alkaline phosphatase activity and the regulation of growth in transformed mammalian cells. J Cell Physiol. 1974 Feb;83(1):27–34. doi: 10.1002/jcp.1040830105. [DOI] [PubMed] [Google Scholar]
- Sibley C. H., Tomkins G. M. Mechanisms of steroid resistance. Cell. 1974 Aug;2(4):221–227. doi: 10.1016/0092-8674(74)90014-2. [DOI] [PubMed] [Google Scholar]
- Symonds G., Sachs L. Activation of normal genes in malignant cells: activation of chemotaxis in relation to other stages of normal differentiation in myeloid leukemia. Somatic Cell Genet. 1979 Nov;5(6):931–944. doi: 10.1007/BF01542652. [DOI] [PubMed] [Google Scholar]