Abstract
Mouse protein 25α (MO25α) is a 40-kDa protein that, together with the STE20-related adaptor-α (STRADα) pseudo kinase, forms a regulatory complex capable of stimulating the activity of the LKB1 tumor suppressor protein kinase. The latter is mutated in the inherited Peutz-Jeghers cancer syndrome (PJS). MO25α binds directly to a conserved Trp-Glu-Phe sequence at the STRADα C terminus, markedly enhancing binding of STRADα to LKB1 and increasing LKB1 catalytic activity. The MO25α crystal structure reveals a helical repeat fold, distantly related to the Armadillo proteins. A complex with the STRADα peptide reveals a hydrophobic pocket that is involved in a unique and specific interaction with the Trp-Glu-Phe motif, further supported by mutagenesis studies. The data represent a first step toward structural analysis of the LKB1–STRAD–MO25 complex, and suggests that MO25α is a scaffold protein to which other regions of STRAD–LKB1, cellular LKB1 substrates or regulatory components could bind.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Buy this article
Purchase on Springer Link
Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Boudeau, J., Sapkota, G. & Alessi, D.R. LKB1, a protein kinase regulating cell proliferation and polarity. FEBS Lett. 546, 159–165 (2003).
Hemminki, A. et al. A serine/threonine kinase gene defective in Peutz-Jeghers syndrome. Nature 391, 184–187 (1998).
Jenne, D.E. et al. Peutz-Jeghers syndrome is caused by mutations in a novel serine threonine kinase. Nat. Genet. 18, 38–44 (1998).
Ylikorkala, A. et al. Vascular abnormalities and deregulation of VEGF in LKB1 deficient mice. Science 293, 1323–1326 (2001).
Bardeesy, N. et al. Loss of the LKB1 tumour suppressor provokes intestinal polyposis but resistance to transformation. Nature 419, 162–167 (2002).
Jishage, K. et al. Role of LKB1, the causative gene of Peutz-Jeghers syndrome, in embryogenesis and polyposis. Proc. Natl. Acad. Sci. USA 99, 8903–8908 (2002).
Miyoshi, H. et al. Gastrointestinal hamartomatous polyposis in LKB1 heterozygous knockout mice. Cancer Res. 62, 2261–2266 (2002).
Nakau, M. et al. Hepatocellular carcinoma caused by loss of heterozygosity in LKB1 gene knockout mice. Cancer Res. 62, 4549–4553 (2002).
Tiainen, M., Ylikorkala, A. & Makela, T.P. Growth suppression by LKB1 is mediated by a G(1) cell cycle arrest. Proc. Natl. Acad. Sci. USA 96, 9248–9251 (1999).
Karuman, P. et al. The Peutz-Jeghers gene product LKB1 is a mediator of p53-dependent cell death. Mol. Cell 7, 1307–1319 (2001).
Tiainen, M., Vaahtomeri, K., Ylikorkala, A. & Makela, T.P. Growth arrest by the LKB1 tumor suppressor: induction of p21(WAF1/CIP1). Hum. Mol. Genet. 11, 1497–1504 (2002).
Watts, J.L., Morton, D.G., Bestman, J. & Kemphues, K.J. The C. elegans par-4 gene encodes a putative serine-threonine kinase required for establishing embryonic asymmetry. Development 127, 1467–1475 (2000).
Martin, S.G. & St. Johnston, D. A role for Drosophila LKB1 in anterior-posterior axis formation and epithelial polarity. Nature 421, 379–384 (2003).
Ossipova, O., Bardeesy, N., DePinho, R.A. & Green, J.B. LKB1 (XEEK1) regulates Wnt signalling in vertebrate. Nat. Cell Biol. 5, 889–894 (2003).
Spicer, J. et al. Regulation of the Wnt signalling component PAR1A by the Peutz-Jeghers syndrome kinase LKB1. Oncogene 22, 4752–4756 (2003).
Baas, A.F. et al. Activation of the tumour suppressor kinase LKB1 by the STE20-like pseudokinase. EMBO J. 22, 3062–3072 (2003).
Boudeau, J. et al. MO25 isoforms interact with STRADα/β enhancing their ability to bind, activate and localise LKB1. EMBO J. 22, 5102–5114 (2003).
Hawley, S.A. et al. Complexes between the LKB1 tumour suppressor, STRADα/β and MO25α/β are upstream kinases in the AMP-activated protein kinase cascade. J. Biol. 2, 28 (2003).
Woods, A. et al. LKB1 is the upstream kinase in the AMP-activated protein kinase cascade. Curr. Biol. 13, 2004–2008 (2003).
Perrakis, A., Morris, R. & Lamzin, V.S. Automated protein model building combined with iterative structure refinement. Nat. Struct. Biol. 6, 458–463 (1999).
Miyamoto, H., Matsushiro, A. & Nozaki, M. Molecular-cloning of a novel messenger-RNA sequence expressed in cleavage stage mouse embryos. Mol. Reprod. Dev. 34, 1–7 (1993).
Nozaki, M., Onishi, Y., Togashi, S. & Miyamoto, H. Molecular characterization of the Drosophila Mo25 gene, which is conserved among Drosophila, mouse, and yeast. DNA Cell Biol. 15, 505–509 (1996).
Chattopadhyaya, R., Meador, W.E., Means, A.R. & Quiocho, F.A. Calmodulin structure refined at 1.7 Å resolution. J. Mol. Biol. 228, 1177–1192 (1992).
Murzin, A.G., Brenner, S.E., Hubbard, T. & Chothia, C. SCOP: a structural classification of proteins database for the investigation of sequences and structures. J. Mol. Biol. 247, 53–54 (1995).
Holm, L. & Sander, C. Protein structure comparison by alignment of distance matrices. J. Mol. Biol. 233, 123–138 (1993).
Kobe, B. Autoinhibition by an internal nuclear localization signal revealed by the crystal structure of mammalian importin α. Nat. Struct. Biol. 6, 388–397 (1999).
Vetter, I.R., Arndt, A., Kutay, U., Gorlich, D. & Wittinghofer, A. Structural view of the Ran-Importin β interaction at 2.3 Å resolution. Cell 97, 635–646 (1999).
Conti, E., Uy, M., Leighton, L., Blobel, G. & Kuriyan, J. Crystallographic analysis of the recognition of a nuclear localization signal by the nuclear import factor karyopherin α. Cell 94, 193–204 (1998).
Huber, A.H., Nelson, W.J. & Weis, W.I. Structural analysis of the armadillo repeat region of β-catenin and its interactions with cadherins. FASEB J. 11, 2510 (1997).
Edwards, T.A., Pyle, S.E., Wharton, R.P. & Aggarwal, A.K. Structure of Pumilio reveals similarity between RNA and peptide binding motifs. Cell 105, 281–289 (2001).
Wang, X.Q., Zamore, P.D. & Hall, T.M.T. Crystal structure of a Pumilio homology domain. Mol. Cell 7, 855–865 (2001).
Wang, X.Q., McLachlan, J., Zamore, P.D. & Hall, T.M.T. Modular recognition of RNA by a human Pumilio-homology domain. Cell 110, 501–512 (2002).
Groves, M.R. & Barford, D. Topological characteristics of helical repeat proteins. Curr. Opin. Struct. Biol. 9, 383–389 (1999).
Andrade, M.A., Perez-Iratxeta, C. & Ponting, C.P. Protein repeats: structures, functions, and evolution. J. Struct. Biol. 134, 117–131 (2001).
Hatzfeld, M. The armadillo family of structural proteins. Int. Rev. Cytol. 186, 179–224 (1999).
Edwards, T.A., Trincao, J., Escalante, C.R., Wharton, R.P. & Aggarwal, A.K. Crystallization and characterization of Pumilio: a novel RNA binding protein. J. Struct. Biol. 132, 251–254 (2000).
Bayliss, R., Littlewood, T., Strawn, L.A., Wente, S.R. & Stewart, M. GLFG and FxFG nucleoporins bind to overlapping sites on importin-β. J. Biol. Chem. 277, 50597–50606 (2002).
Cingolani, G., Bednenko, J., Gillespie, M.T. & Gerace, L. Molecular basis for the recognition of a nonclassical nuclear localization signal by importin β. Mol. Cell 10, 1345–1353 (2002).
Conti, E. & Kuriyan, J. Crystallographic analysis of the specific yet versatile recognition of distinct nuclear localization signals by karyopherin α. Struct. Fold. Des. 8, 329–338 (2000).
Graham, T.A., Weaver, C., Mao, F., Kimelman, D. & Xu, W.Q. Crystal structure of a β-catenin/Tcf complex. Cell 103, 885–896 (2000).
Huber, A.H. & Weis, W.I. The structure of the β-catenin/E-cadherin complex and the molecular basis of diverse ligand recognition by β-catenin. Cell 105, 391–402 (2001).
Daniels, D.L. & Weis, W.I. ICAT inhibits β-catenin binding to Tcf/Lef-family transcription factors and the general coactivator p300 using independent structural modules. Mol. Cell 10, 573–584 (2002).
Alphey, M.S. et al. The high resolution crystal structure of recombinant Crithidia fasciculata tryparedoxin-I. J. Biol. Chem. 274, 25613–25622 (1999).
Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).
Terwilliger, T.C. & Berendzen, J. Automated MAD and MIR structure solution. Acta Crystallogr. D 55, 849–861 (1999).
Cowtan, K. Joint CCP4 and ESF-EACBM Newsletter on Protein Crystallography Vol. 31 (Daresbury Laboratory, Warrington, UK, 1994).
Jones, T.A., Zou, J.Y., Cowan, S.W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A 47, 110–119 (1991).
Brunger, A.T. et al. Crystallography and NMR system: a new software system for macromolecular structure determination. Acta Crystallogr. D 54, 905–921 (1998).
Navaza, J. AMoRe: an automated package for molecular replacement. Acta Crystallogr. A 50, 157–163 (1994).
Murshudov, G.N., Vagin, A.A. & Dodson, E.J. Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr. D 53, 240–255 (1997).
Winn, M.D., Isupov, M.N. & Murshudov, G.N. Use of TLS parameters to model anisotropic displacements in macromolecular refinement. Acta Crystallogr. D 57, 122–133 (2001).
Kleywegt, G.J., Zou, J.Y., Kjeldgaard, M. & Jones, T.A., Around, O. In International Tables for Crystallography Vol. F (eds. Rossman, M.G. & Arnold, E.) Ch. 17.1 353–356, 366–367 Kluwer Academic, Dordrecht, The Netherlands, 2000).
Nicholls, A., Sharp, K. & Honig, B. Protein folding and association—insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins 11, 281–296 (1991).
Acknowledgements
We thank the European Synchrotron Radiation Facility (Grenoble, France) for the time at beamlines ID14-EH4. C.C.M. is supported by a Biotechnology and Biological Sciences Research Council CASE studentship, D.M.F.v.A. by a Wellcome Trust Career Development Research Fellowship and an European Molecular Biology Organization Young Investigator Fellowship, D.R.A. by the Medical Research Council (UK), Diabetes UK, Association for International Cancer Research. D.M.F.v.A. and D.R.A. are also supported by the pharmaceutical companies supporting the Division of Signal Transduction Therapy unit in Dundee (AstraZeneca, Boehringer-Ingelheim, GlaxoSmithKline, Merck & Co. Inc, Merck KGA and Pfizer).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Rights and permissions
About this article
Cite this article
Milburn, C., Boudeau, J., Deak, M. et al. Crystal structure of MO25α in complex with the C terminus of the pseudo kinase STE20-related adaptor. Nat Struct Mol Biol 11, 193–200 (2004). https://doi.org/10.1038/nsmb716
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nsmb716
This article is cited by
-
RBM4 dictates ESCC cell fate switch from cellular senescence to glutamine-addiction survival through inhibiting LKB1-AMPK-axis
Signal Transduction and Targeted Therapy (2023)
-
Lanthanum Chloride Induces Axon Abnormality Through LKB1-MARK2 and LKB1-STK25-GM130 Signaling Pathways
Cellular and Molecular Neurobiology (2023)
-
MicroRNA-22 promoted osteogenic differentiation of valvular interstitial cells by inhibiting CAB39 expression during aortic valve calcification
Cellular and Molecular Life Sciences (2022)
-
Germinal center kinases in immune regulation
Cellular & Molecular Immunology (2012)
-
MO25 is a master regulator of SPAK/OSR1 and MST3/MST4/YSK1 protein kinases
The EMBO Journal (2011)