Phosphorylation Disrupts the Central Helix in Op18/stathmin and Suppresses Binding to Tubulin

Overview

Journal EMBO Rep

Specialty Molecular Biology

Date 2001 Jun 21

PMID 11415983

Citations 13

Authors

M O Steinmetz

W Jahnke

H Towbin

C Garcia-Echeverria

H Voshol

D Muller

J van Oostrum

Affiliations

Soon will be listed here.

Abstract

Protein phosphorylation represents a ubiquitous control mechanism in living cells. The structural prerequisites and consequences of this important post-translational modification, however, are poorly understood. Oncoprotein 18/stathmin (Op18) is a globally disordered phosphoprotein that is involved in the regulation of the microtubule (MT) filament system. Here we document that phosphorylation of Ser63, which is located within a helix initiation site in Op18, disrupts the transiently formed amphipathic helix. The phosphoryl group reduces tubulin binding 10-fold and suppresses the MT polymerization inhibition activity of Op18's C-terminal domain. Op18 represents an example where phosphorylation occurs within a regular secondary structural element. Together, our findings have implications for the prediction of phosphorylation sites and give insights into the molecular behavior of a globally disordered protein.

Citing Articles

Tubulin Regulates the Stability and Localization of STMN2 by Binding Preferentially to Its Soluble Form.

Deng X, Bradshaw G, Kalocsay M, Mitchison T bioRxiv. 2025; .

PMID: 40060442 PMC: 11888388. DOI: 10.1101/2025.02.27.640326.

The Origin of Discrepancies between Predictions and Annotations in Intrinsically Disordered Proteins.

Pajkos M, Erdos G, Dosztanyi Z Biomolecules. 2023; 13(10).

PMID: 37892124 PMC: 10604070. DOI: 10.3390/biom13101442.

Stathmins and Motor Neuron Diseases: Pathophysiology and Therapeutic Targets.

Gagliardi D, Pagliari E, Meneri M, Melzi V, Rizzo F, Comi G Biomedicines. 2022; 10(3).

PMID: 35327513 PMC: 8945549. DOI: 10.3390/biomedicines10030711.

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.

The Cytoskeleton-A Complex Interacting Meshwork.

Hohmann T, Dehghani F Cells. 2019; 8(4).

PMID: 31003495 PMC: 6523135. DOI: 10.3390/cells8040362.

References

Larsson N, Segerman B, Howell B, Fridell K, Cassimeris L, Gullberg M . Op18/stathmin mediates multiple region-specific tubulin and microtubule-regulating activities. J Cell Biol. 1999; 146(6):1289-302. PMC: 2156119. DOI: 10.1083/jcb.146.6.1289. View

Muller D, Schindler P, COULOT M, Voshol H, Oostrum J . Mass spectrometric characterization of stathmin isoforms separated by 2D PAGE. J Mass Spectrom. 1999; 34(4):336-45. DOI: 10.1002/(SICI)1096-9888(199904)34:4<336::AID-JMS765>3.0.CO;2-U. View

WALLON G, Rappsilber J, Mann M, Serrano L . Model for stathmin/OP18 binding to tubulin. EMBO J. 2000; 19(2):213-22. PMC: 305555. DOI: 10.1093/emboj/19.2.213. View

Steinmetz M, Kammerer R, Jahnke W, Goldie K, Lustig A, Oostrum J . Op18/stathmin caps a kinked protofilament-like tubulin tetramer. EMBO J. 2000; 19(4):572-80. PMC: 305595. DOI: 10.1093/emboj/19.4.572. View

Ostedgaard L, Baldursson O, Vermeer D, Welsh M, Robertson A . A functional R domain from cystic fibrosis transmembrane conductance regulator is predominantly unstructured in solution. Proc Natl Acad Sci U S A. 2000; 97(10):5657-62. PMC: 25884. DOI: 10.1073/pnas.100588797. View

Daly N, Hoffmann R, Otvos Jr L, Craik D . Role of phosphorylation in the conformation of tau peptides implicated in Alzheimer's disease. Biochemistry. 2000; 39(30):9039-46. DOI: 10.1021/bi0004807. View

Gigant B, Curmi P, Martin-Barbey C, Charbaut E, Lachkar S, Lebeau L . The 4 A X-ray structure of a tubulin:stathmin-like domain complex. Cell. 2000; 102(6):809-16. DOI: 10.1016/s0092-8674(00)00069-6. View

Lupas A, Van Dyke M, Stock J . Predicting coiled coils from protein sequences. Science. 1991; 252(5009):1162-4. DOI: 10.1126/science.252.5009.1162. View

Sobel A . Stathmin: a relay phosphoprotein for multiple signal transduction?. Trends Biochem Sci. 1991; 16(8):301-5. DOI: 10.1016/0968-0004(91)90123-d. View

10.

Schweers O, Marx A, Mandelkow E . Structural studies of tau protein and Alzheimer paired helical filaments show no evidence for beta-structure. J Biol Chem. 1994; 269(39):24290-7. View

11.

Smith L, Bolin K, Schwalbe H, MacArthur M, Thornton J, Dobson C . Analysis of main chain torsion angles in proteins: prediction of NMR coupling constants for native and random coil conformations. J Mol Biol. 1996; 255(3):494-506. DOI: 10.1006/jmbi.1996.0041. View

12.

Belmont L, Mitchison T . Identification of a protein that interacts with tubulin dimers and increases the catastrophe rate of microtubules. Cell. 1996; 84(4):623-31. DOI: 10.1016/s0092-8674(00)81037-5. View

13.

Szilak L, Moitra J, Krylov D, Vinson C . Phosphorylation destabilizes alpha-helices. Nat Struct Biol. 1997; 4(2):112-4. DOI: 10.1038/nsb0297-112. View

14.

Larsson N, Marklund U, Gradin H, Brattsand G, Gullberg M . Control of microtubule dynamics by oncoprotein 18: dissection of the regulatory role of multisite phosphorylation during mitosis. Mol Cell Biol. 1997; 17(9):5530-9. PMC: 232401. DOI: 10.1128/MCB.17.9.5530. View

15.

Curmi P, Andersen S, Lachkar S, Gavet O, Karsenti E, Knossow M . The stathmin/tubulin interaction in vitro. J Biol Chem. 1997; 272(40):25029-36. DOI: 10.1074/jbc.272.40.25029. View

16.

Smith J, Scholtz J . Energetics of polar side-chain interactions in helical peptides: salt effects on ion pairs and hydrogen bonds. Biochemistry. 1998; 37(1):33-40. DOI: 10.1021/bi972026h. View

17.

Lawler S . Microtubule dynamics: if you need a shrink try stathmin/Op18. Curr Biol. 1998; 8(6):R212-4. DOI: 10.1016/s0960-9822(98)70128-9. View

18.

Johnson L, Lowe E, Noble M, Owen D . The Eleventh Datta Lecture. The structural basis for substrate recognition and control by protein kinases. FEBS Lett. 1998; 430(1-2):1-11. DOI: 10.1016/s0014-5793(98)00606-1. View

19.

Kreegipuu A, Blom N, Brunak S, Jarv J . Statistical analysis of protein kinase specificity determinants. FEBS Lett. 1998; 430(1-2):45-50. DOI: 10.1016/s0014-5793(98)00503-1. View

20.

Tholey A, Lindemann A, Kinzel V, Reed J . Direct effects of phosphorylation on the preferred backbone conformation of peptides: a nuclear magnetic resonance study. Biophys J. 1999; 76(1 Pt 1):76-87. PMC: 1302501. DOI: 10.1016/S0006-3495(99)77179-1. View