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. 1992 Oct 15;287(Pt 2):549–554. doi: 10.1042/bj2870549

Expression of transfected stathmin cDNA reveals novel phosphorylated forms associated with developmental and functional cell regulation.

V Doye 1, S Le Gouvello 1, T Dobransky 1, H Chneiweiss 1, L Beretta 1, A Sobel 1
PMCID: PMC1133199  PMID: 1445213

Abstract

Stathmin is a ubiquitous, highly conserved phosphoprotein, which most likely acts as an intracellular relay integrating various transduction pathways triggered by extracellular signals. Two post-translational isoforms (alpha and beta) have been previously identified whose increasingly phosphorylated forms migrate as a set of isoelectric variant spots (molecular mass 19 kDa; pI 6.2-5.6) on two-dimensional electrophoretic gels. In parallel with the phosphorylation of these forms of stathmin, two sets of three proteins migrating with slightly higher apparent molecular masses (21 and 23 kDa respectively) also incorporated radioactive phosphate in response to cell regulation through various transduction pathways. These phosphoproteins, previously referred to as proteins '16' and '17', share several biochemical properties with stathmin and are recognized by antibodies directed to stathmin or to stathmin peptides. Furthermore, when rat stathmin cDNA was transfected into mouse myogenic C2 cells, it directed the expression of protein sets 16 and 17 together with the 19 kDa forms of stathmin, as detected with a species-specific anti-stathmin antiserum. Proteins 16 and 17 are thus novel phosphorylated derivatives of stathmin, encoded by the same cDNA as its previously identified 19 kDa forms. These results increase the known complexity and diversity of stathmin patterns, which may yield the molecular support for its proposed role as a relay integrating various signals which regulate the proliferation, differentiation and functions of cells during development and adult life.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beretta L., Boutterin M. C., Sobel A. Phosphorylation of intracellular proteins related to the multihormonal regulation of prolactin: comparison of normal anterior pituitary cells in culture with the tumor-derived GH cell lines. Endocrinology. 1988 Jan;122(1):40–51. doi: 10.1210/endo-122-1-40. [DOI] [PubMed] [Google Scholar]
  2. Beretta L., Houdouin F., Sobel A. Identification of two distinct isoforms of stathmin and characterization of their respective phosphorylated forms. J Biol Chem. 1989 Jun 15;264(17):9932–9938. [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Braverman R., Bhattacharya B., Feuerstein N., Cooper H. L. Identification and characterization of the nonphosphorylated precursor of pp17, a phosphoprotein associated with phorbol ester induction of growth arrest and monocytic differentiation in HL-60 promyelocytic leukemia cells. J Biol Chem. 1986 Oct 25;261(30):14342–14348. [PubMed] [Google Scholar]
  5. Chneiweiss H., Beretta L., Cordier J., Boutterin M. C., Glowinski J., Sobel A. Stathmin is a major phosphoprotein and cyclic AMP-dependent protein kinase substrate in mouse brain neurons but not in astrocytes in culture: regulation during ontogenesis. J Neurochem. 1989 Sep;53(3):856–863. doi: 10.1111/j.1471-4159.1989.tb11783.x. [DOI] [PubMed] [Google Scholar]
  6. Chneiweiss H., Cordier J., Sobel A. Stathmin phosphorylation is regulated in striatal neurons by vasoactive intestinal peptide and monoamines via multiple intracellular pathways. J Neurochem. 1992 Jan;58(1):282–289. doi: 10.1111/j.1471-4159.1992.tb09308.x. [DOI] [PubMed] [Google Scholar]
  7. Cooper H. L., Fuldner R., McDuffie E., Braverman R. A specific defect of prosolin phosphorylation in T cell leukemic lymphoblasts is associated with impaired down-regulation of DNA synthesis. J Immunol. 1990 Aug 15;145(4):1205–1213. [PubMed] [Google Scholar]
  8. Cooper H. L., Fuldner R., McDuffie E., Braverman R. T cell receptor activation induces rapid phosphorylation of prosolin, which mediates down-regulation of DNA synthesis in proliferating peripheral lymphocytes. J Immunol. 1991 Jun 1;146(11):3689–3696. [PubMed] [Google Scholar]
  9. Cooper H. L., McDuffie E., Braverman R. Human peripheral lymphocyte growth regulation and response to phorbol esters is linked to synthesis and phosphorylation of the cytosolic protein, prosolin. J Immunol. 1989 Aug 1;143(3):956–963. [PubMed] [Google Scholar]
  10. Doye V., Boutterin M. C., Sobel A. Phosphorylation of stathmin and other proteins related to nerve growth factor-induced regulation of PC12 cells. J Biol Chem. 1990 Jul 15;265(20):11650–11655. [PubMed] [Google Scholar]
  11. Doye V., Kellermann O., Buc-Caron M. H., Sobel A. High expression of stathmin in multipotential teratocarcinoma and normal embryonic cells versus their early differentiated derivatives. Differentiation. 1992 Jun;50(2):89–96. doi: 10.1111/j.1432-0436.1992.tb00489.x. [DOI] [PubMed] [Google Scholar]
  12. Doye V., Soubrier F., Bauw G., Boutterin M. C., Beretta L., Koppel J., Vandekerckhove J., Sobel A. A single cDNA encodes two isoforms of stathmin, a developmentally regulated neuron-enriched phosphoprotein. J Biol Chem. 1989 Jul 25;264(21):12134–12137. [PubMed] [Google Scholar]
  13. Feuerstein N., Nishikawa M., Cooper H. L. Cell-free system studies on the phosphorylation of the 17,000-20,000 dalton protein induced by phorbol ester in human leukemic cells and evidence for a similar event in virally transformed murine fibroblasts. Cancer Res. 1985 Jul;45(7):3243–3251. [PubMed] [Google Scholar]
  14. Gullberg M., Noreus K., Brattsand G., Friedrich B., Shingler V. Purification and characterization of a 19-kilodalton intracellular protein. An activation-regulated putative protein kinase C substrate of T lymphocytes. J Biol Chem. 1990 Oct 15;265(29):17499–17505. [PubMed] [Google Scholar]
  15. Hailat N., Strahler J., Melhem R., Zhu X. X., Brodeur G., Seeger R. C., Reynolds C. P., Hanash S. N-myc gene amplification in neuroblastoma is associated with altered phosphorylation of a proliferation related polypeptide (Op18). Oncogene. 1990 Nov;5(11):1615–1618. [PubMed] [Google Scholar]
  16. Hanash S. M., Strahler J. R., Kuick R., Chu E. H., Nichols D. Identification of a polypeptide associated with the malignant phenotype in acute leukemia. J Biol Chem. 1988 Sep 15;263(26):12813–12815. [PubMed] [Google Scholar]
  17. Hunter T. A thousand and one protein kinases. Cell. 1987 Sep 11;50(6):823–829. doi: 10.1016/0092-8674(87)90509-5. [DOI] [PubMed] [Google Scholar]
  18. Koppel J., Boutterin M. C., Doye V., Peyro-Saint-Paul H., Sobel A. Developmental tissue expression and phylogenetic conservation of stathmin, a phosphoprotein associated with cell regulations. J Biol Chem. 1990 Mar 5;265(7):3703–3707. [PubMed] [Google Scholar]
  19. 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]
  20. Maucuer A., Doye V., Sobel A. A single amino acid difference distinguishes the human and the rat sequences of stathmin, a ubiquitous intracellular phosphoprotein associated with cell regulations. FEBS Lett. 1990 May 21;264(2):275–278. doi: 10.1016/0014-5793(90)80266-l. [DOI] [PubMed] [Google Scholar]
  21. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  22. Pasmantier R., Danoff A., Fleischer N., Schubart U. K. P19, a hormonally regulated phosphoprotein of peptide hormone-producing cells: secretagogue-induced phosphorylation in AtT-20 mouse pituitary tumor cells and in rat and hamster insulinoma cells. Endocrinology. 1986 Sep;119(3):1229–1238. doi: 10.1210/endo-119-3-1229. [DOI] [PubMed] [Google Scholar]
  23. Peyron J. F., Aussel C., Ferrua B., Häring H., Fehlmann M. Phosphorylation of two cytosolic proteins. An early event of T-cell activation. Biochem J. 1989 Mar 1;258(2):505–510. doi: 10.1042/bj2580505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roach P. J. Multisite and hierarchal protein phosphorylation. J Biol Chem. 1991 Aug 5;266(22):14139–14142. [PubMed] [Google Scholar]
  25. Schubart U. K., Banerjee M. D., Eng J. Homology between the cDNAs encoding phosphoprotein p19 and SCG10 reveals a novel mammalian gene family preferentially expressed in developing brain. DNA. 1989 Jul-Aug;8(6):389–398. doi: 10.1089/dna.1.1989.8.389. [DOI] [PubMed] [Google Scholar]
  26. Schubart U. K. Expression of phosphoprotein p19 in brain, testis, and neuroendocrine tumor cells. Developmental regulation in rat brain. J Biol Chem. 1988 Aug 25;263(24):12156–12160. [PubMed] [Google Scholar]
  27. Sobel A., Boutterin M. C., Beretta L., Chneiweiss H., Doye V., Peyro-Saint-Paul H. Intracellular substrates for extracellular signaling. Characterization of a ubiquitous, neuron-enriched phosphoprotein (stathmin). J Biol Chem. 1989 Mar 5;264(7):3765–3772. [PubMed] [Google Scholar]
  28. Sobel A. Stathmin: a relay phosphoprotein for multiple signal transduction? Trends Biochem Sci. 1991 Aug;16(8):301–305. doi: 10.1016/0968-0004(91)90123-d. [DOI] [PubMed] [Google Scholar]
  29. Sobel A., Tashjian A. H., Jr Distinct patterns of cytoplasmic protein phosphorylation related to regulation of synthesis and release of prolactin by GH cells. J Biol Chem. 1983 Sep 10;258(17):10312–10324. [PubMed] [Google Scholar]
  30. Zhu X. X., Kozarsky K., Strahler J. R., Eckerskorn C., Lottspeich F., Melhem R., Lowe J., Fox D. A., Hanash S. M., Atweh G. F. Molecular cloning of a novel human leukemia-associated gene. Evidence of conservation in animal species. J Biol Chem. 1989 Aug 25;264(24):14556–14560. [PubMed] [Google Scholar]
  31. le Gouvello S., Chneiweiss H., Tarantino N., Debre P., Sobel A. Stathmin phosphorylation patterns discriminate between distinct transduction pathways of human T lymphocyte activation through CD2 triggering. FEBS Lett. 1991 Aug 5;287(1-2):80–84. doi: 10.1016/0014-5793(91)80020-4. [DOI] [PubMed] [Google Scholar]

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