Regulation of Microtubule Dynamics by Extracellular Signals: CAMP-dependent Protein Kinase Switches off the Activity of Oncoprotein 18 in Intact Cells

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 1998 Feb 14

PMID 9425161

Citations 33

Authors

H M Gradin

N Larsson

U Marklund

M Gullberg

Affiliations

Soon will be listed here.

Abstract

Oncoprotein 18 (Op18, also termed p19, 19K, metablastin, stathmin, and prosolin) is a recently identified regulator of microtubule (MT) dynamics. Op18 is a target for both cell cycle and cell surface receptor-coupled kinase systems, and phosphorylation of Op18 on specific combinations of sites has been shown to switch off its MT-destabilizing activity. Here we show that induced expression of the catalytic subunit of cAMP-dependent protein kinase (PKA) results in a dramatic increase in cellular MT polymer content concomitant with phosphorylation and partial degradation of Op18. That PKA may regulate the MT system by downregulation of Op18 activity was evaluated by a genetic system allowing conditional co-expression of PKA and a series of kinase target site-deficient mutants of Op18. The results show that phosphorylation of Op18 on two specific sites, Ser-16 and Ser-63, is necessary and sufficient for PKA to switch off Op18 activity in intact cells. The regulatory importance of dual phosphorylation on Ser-16 and Ser-63 of Op18 was reproduced by in vitro assays. These results suggest a simple model where PKA phosphorylation downregulates the MT-destabilizing activity of Op18, which in turn promotes increased tubulin polymerization. Hence, the present study shows that Op18 has the potential to regulate the MT system in response to external signals such as cAMP-linked agonists.

Citing Articles

Molecular interactions at the colchicine binding site in tubulin: An X-ray crystallography perspective.

Wang J, Miller D, Li W Drug Discov Today. 2021; 27(3):759-776.

PMID: 34890803 PMC: 8901563. DOI: 10.1016/j.drudis.2021.12.001.

Stathmin dynamics modulate the activity of eribulin in breast cancer cells.

Yoshie M, Ishida A, Ohashi H, Nakachi N, Azumi M, Tamura K Pharmacol Res Perspect. 2021; 9(4):e00786.

PMID: 34176226 PMC: 8236080. DOI: 10.1002/prp2.786.

Stathmin Regulates Spatiotemporal Variation in the Memory Loop in Single-Prolonged Stress Rats.

Shan W, Han F, Xu Y, Shi Y J Mol Neurosci. 2020; 70(4):576-589.

PMID: 31933182 DOI: 10.1007/s12031-019-01459-w.

SIAH ubiquitin ligases regulate breast cancer cell migration and invasion independent of the oxygen status.

Adam M, Matt S, Christian S, Hess-Stumpp H, Haegebarth A, Hofmann T Cell Cycle. 2015; 14(23):3734-47.

PMID: 26654769 PMC: 4825722. DOI: 10.1080/15384101.2015.1104441.

PIWIL1 destabilizes microtubule by suppressing phosphorylation at Ser16 and RLIM-mediated degradation of Stathmin1.

Li C, Zhou X, Chen J, Lu Y, Sun Q, Tao D Oncotarget. 2015; 6(29):27794-804.

PMID: 26317901 PMC: 4695026. DOI: 10.18632/oncotarget.4533.

References

Strahler J, Hailat N, LAMB B, Rogers K, Underhill J, Melhem R . Activation of resting peripheral blood lymphocytes through the T cell receptor induces rapid phosphorylation of Op18. J Immunol. 1992; 149(4):1191-8. View

Horwitz S, Shen H, He L, Dittmar P, Neef R, Chen J . The microtubule-destabilizing activity of metablastin (p19) is controlled by phosphorylation. J Biol Chem. 1997; 272(13):8129-32. DOI: 10.1074/jbc.272.13.8129. View

Ball R, Albrecht T, Thompson W, James O, Carney D . Thrombin, epidermal growth factor, and phorbol myristate acetate stimulate tubulin polymerization in quiescent cells: a potential link to mitogenesis. Cell Motil Cytoskeleton. 1992; 23(4):265-78. DOI: 10.1002/cm.970230406. View

Mitchison T . Compare and contrast actin filaments and microtubules. Mol Biol Cell. 1992; 3(12):1309-15. PMC: 275701. DOI: 10.1091/mbc.3.12.1309. View

Brattsand G, Roos G, Marklund U, Ueda H, Landberg G, Nanberg E . Quantitative analysis of the expression and regulation of an activation-regulated phosphoprotein (oncoprotein 18) in normal and neoplastic cells. Leukemia. 1993; 7(4):569-79. View

Wang Y, Liao P, Allison J, Gage D, Andrews P, Lubman D . Phorbol 12-myristate 13-acetate-induced phosphorylation of Op18 in Jurkat T cells. Identification of phosphorylation sites by matrix-assisted laser desorption ionization mass spectrometry. J Biol Chem. 1993; 268(19):14269-77. View

Marklund U, Brattsand G, Shingler V, Gullberg M . Serine 25 of oncoprotein 18 is a major cytosolic target for the mitogen-activated protein kinase. J Biol Chem. 1993; 268(20):15039-47. View

Beretta L, Dobransky T, Sobel A . Multiple phosphorylation of stathmin. Identification of four sites phosphorylated in intact cells and in vitro by cyclic AMP-dependent protein kinase and p34cdc2. J Biol Chem. 1993; 268(27):20076-84. View

Roos G, Brattsand G, Landberg G, Marklund U, Gullberg M . Expression of oncoprotein 18 in human leukemias and lymphomas. Leukemia. 1993; 7(10):1538-46. View

10.

Marklund U, Brattsand G, Osterman O, Ohlsson P, Gullberg M . Multiple signal transduction pathways induce phosphorylation of serines 16, 25, and 38 of oncoprotein 18 in T lymphocytes. J Biol Chem. 1993; 268(34):25671-80. View

11.

Brattsand G, Marklund U, Nylander K, Roos G, Gullberg M . Cell-cycle-regulated phosphorylation of oncoprotein 18 on Ser16, Ser25 and Ser38. Eur J Biochem. 1994; 220(2):359-68. DOI: 10.1111/j.1432-1033.1994.tb18632.x. View

12.

Cantrell D . G proteins in lymphocyte signalling. Curr Opin Immunol. 1994; 6(3):380-4. DOI: 10.1016/0952-7915(94)90116-3. View

13.

Marklund U, Osterman O, Melander H, Bergh A, Gullberg M . The phenotype of a "Cdc2 kinase target site-deficient" mutant of oncoprotein 18 reveals a role of this protein in cell cycle control. J Biol Chem. 1994; 269(48):30626-35. View

14.

Jameson L, CAPLOW M . Modification of microtubule steady-state dynamics by phosphorylation of the microtubule-associated proteins. Proc Natl Acad Sci U S A. 1981; 78(6):3413-7. PMC: 319578. DOI: 10.1073/pnas.78.6.3413. View

15.

Brattin Jr W, Portanova R . Dibutyryl cyclic AMP-induced phosphorylation of specific proteins in adenohypophysial cells. Mol Cell Endocrinol. 1981; 23(1):77-90. DOI: 10.1016/0303-7207(81)90118-0. View

16.

Ookata K, Hisanaga S, Bulinski J, Murofushi H, Aizawa H, Itoh T . Cyclin B interaction with microtubule-associated protein 4 (MAP4) targets p34cdc2 kinase to microtubules and is a potential regulator of M-phase microtubule dynamics. J Cell Biol. 1995; 128(5):849-62. PMC: 2120387. DOI: 10.1083/jcb.128.5.849. View

17.

Cleveland D, Lopata M, Sherline P, Kirschner M . Unpolymerized tubulin modulates the level of tubulin mRNAs. Cell. 1981; 25(2):537-46. DOI: 10.1016/0092-8674(81)90072-6. View

18.

Schubart U . Regulation of protein phosphorylation in hamster insulinoma cells. Identification of Ca2+-regulated cytoskeletal and cAMP-regulated cytosolic phosphoproteins by two-dimensional electrophoresis. J Biol Chem. 1982; 257(20):12231-8. View

19.

Sobel A, TASHJIAN Jr A . Distinct patterns of cytoplasmic protein phosphorylation related to regulation of synthesis and release of prolactin by GH cells. J Biol Chem. 1983; 258(17):10312-24. View

20.

Cole N, Lippincott-Schwartz J . Organization of organelles and membrane traffic by microtubules. Curr Opin Cell Biol. 1995; 7(1):55-64. DOI: 10.1016/0955-0674(95)80045-x. View