Amino Acid Motifs; Amino Acid Sequence; Animals; Binding Sites; Crystallography, X-Ray; Models, Molecular; Molecular Sequence Data; Myosin Heavy Chains/chemistry/metabolism; Myosin Light Chains/*chemistry/metabolism; Myosin Type V/chemistry/metabolism; Protein Binding; *Protein Conformation; Saccharomyces cerevisiae Proteins/*chemistry/metabolism; Sequence Alignment; Support, U.S. Gov't, Non-P.H.S.; Support, U.S. Gov't, P.H.S.
Abstract :
[en] IQ motifs are widespread in nature. Mlc1p is a calmodulin-like myosin light chain that binds to IQ motifs of a class V myosin, Myo2p, and an IQGAP-related protein, Iqg1p, playing a role in polarized growth and cytokinesis in Saccharomyces cerevisiae. The crystal structures of Mlc1p bound to IQ2 and IQ4 of Myo2p differ dramatically. When bound to IQ2, Mlc1p adopts a compact conformation in which both the N- and C-lobes interact with the IQ motif. However, in the complex with IQ4, the N-lobe no longer interacts with the IQ motif, resulting in an extended conformation of Mlc1p. The two light chain structures relate to two distinct subfamilies of IQ motifs, one of which does not interact with the N-lobes of calmodulin-like light chains. The correlation between light chain structure and IQ sequence is demonstrated further by sedimentation velocity analysis of complexes of Mlc1p with IQ motifs from Myo2p and Iqg1p. The resulting 'free' N-lobes of myosin light chains in the extended conformation could mediate the formation of ternary complexes during protein localization and/or partner recruitment.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Terrak, Mohammed ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Wu, G.
Stafford, W. F.
Lu, R. C.
Dominguez, R.
Language :
English
Title :
Two distinct myosin light chain structures are induced by specific variations within the bound IQ motifs-functional implications
Bahler, M. and Rhoads, A. (2002) Calmodulin signaling via the IQ motif. FEBS Lett., 513, 107-113.
Boyne, J.R., Yosuf, H.M., Bieganowski, P., Brenner, C. and Price, C. (2000) Yeast myosin light chain, Mlclp, interacts with both IQGAP and class II myosin to effect cytokinesis. J. Cell Sci., 113, 4533-4543.
Brockerhoff, S.E., Stevens, R.C, and Davis, T.N. (1994) The unconventional myosin, Myo2p, is a calmodulin target at sites of cell growth in Saccharomyces cerevisiae. J. Cell Biol., 124, 315-323.
CCP4 (1994) The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D, 50, 760-763.
Chakrabartty, A., Schellman, J.A. and Baldwin, R.L. (1991) Large differences in the helix propensities of alanine and glycine. Nature, 351, 586-588.
Cheney, R.E., O'Shea, M.K., Heuser, J.E., Coelho, M.V., Wolenski, J.S., Espreafico, E.M., Forscher, P., Larson, R.E. and Mooseker, M.S. (1993) Brain myosin-V is a two-headed unconventional myosin with motor activity. Cell, 75, 13-23.
Cowtan, K. and Main, P. (1998) Miscellaneous algorithms for density modification. Acta Crystallogr. D, 54, 487-493.
Dominguez, R., Freyzon, Y., Trybus, K.M. and Cohen, C. (1998) Crystal structure of a vertebrate smooth muscle myosin motor domain and its complex with the essential light chain: visualization of the pre-power stroke state. Cell, 94, 559-571.
D'Souza, V.M., Naqvi, N.I., Wang, H. and Balasubramanian, M.K. (2001) Interactions of Cdc4p, a myosin light chain, with IQ-domain containing proteins in Schizosaccharomyces pombe. Cell Struct. Funct., 26, 555-565.
Eng, K., Naqvi, N.I., Wong, K.C. and Balasubramanian, M.K. (1998) Rng2p, a protein required for cytokinesis in fission yeast, is a component of the actomyosin ring and the spindle pole body. Curr. Biol., 8, 611-621.
Epp, J.A. and Chant, J. (1997) An IQGAP-related protein controls actinring formation and cytokinesis in yeast. Curr. Biol., 7, 921-929.
GarciadelaTorre, J., Navarro, S., Lopez Martinez, M.C., Diaz, F.G. and Lopez Cascales, J.J. (1994) HYDRO: a computer program for the prediction of hydrodynamic properties of macromolecules. Biophys. J., 67, 530-531.
Govindan, B., Bowser, R. and Novick, P. (1995) The role of Myo2, a yeast class V myosin, in vesicular transport. J. Cell Biol., 128, 1055-1068.
Higgins, D.G., Thompson, J.D. and Gibson, T.J. (1996) Using CLUSTAL for multiple sequence alignments. Methods Enzymol., 266, 383-402.
Houdusse, A. and Cohen, C. (1996) Structure of the regulatory domain of scallop myosin at 2 Å resolution: implications for regulation. Structure, 4, 21-32.
Johnston, G.C., Prendergast, J.A. and Singer, R.A (1991) The Saccharomyces cerevisiae MYO2 gene encodes an essential myosin for vectorial transport of vesicles. J. Cell Biol., 113, 539-551.
Lillie, S.H. and Brown, S.S. (1994) Immunofluorescence localization of the unconventional myosin, Myo2p and the putative kinesin-related protein, Smy1p, to the same regions of polarized growth in Saccharomyces cerevisiae. J. Cell Biol., 125, 825-842.
Lippincott, J. and Li, R. (1998) Sequential assembly of myosin II, an IQGAP-like protein and filamentous actin to a ring structure involved in budding yeast cytokinesis. J. Cell Biol., 140, 355-366.
McCollum, D., Balasubramanian, M.K., Pelcher, L.E., Hemmingsen, S.M. and Gould, K.L. (1995) Schizosaccharomyces pombe cdc4+ gene encodes a novel EF-hand protein essential for cytokinesis. J. Cell Biol., 130, 651-660.
McCollum, D., Feoktistova, A. and Gould, K.L. (1999) Phosphorylation of the myosin-II light chain does not regulate the timing of cytokinesis in fission yeast. J. Biol. Chem., 274, 17691-17695.
Murshudov, G.N., Lebedev, A., Vagin, A.A., Wilson, K.S. and Dodson, E.J. (1999) Efficient anisotropic refinement of macromolecular structures using FFT. Acta Crystallogr. D, 55, 247-255.
Naqvi, N.I., Eng, K., Gould, K.L. and Balasubramanian, M.K. (1999) Evidence for F-actin-dependent and -independent mechanisms involved in assembly and stability of the medial actomyosin ring in fission yeast. EMBO J., 18, 854-862.
Osman, M.A. and Cerione, R.A. (1998) Iqg1p, a yeast homologue of the mammalian IQGAPs, mediates cdc42p effects on the actin cytoskeleton. J. Cell Biol., 142, 443-455.
Pruyne, D.W., Schott, D.H. and Bretscher, A. (1998) Tropomyosin-containing actin cables direct the Myo2p-dependent polarized delivery of secretory vesicles in budding yeast. J. Cell Biol., 143, 1931-1945.
Rayment, I., Rypniewski, W.R., Schmidt-Base, K., Smith, R., Tomchick, D.R., Benning, M.M., Winkelmann, D.A., Wesenberg, G. and Holden, H.M. (1993) Three-dimensional structure of myosin subfragment-1: a molecular motor. Science, 261, 50-58.
Reck-Peterson, S.L., Provance, D.W.,Jr, Mooseker, M.S. and Mercer, J.A. (2000) Class V myosins. Biochim. Biophys. Acta, 1496, 36-51.
Schumacher, M.A., Rivard, A.F., Bachinger, H.P. and Adelman, J.P. (2001) Structure of the gating domain of a Ca2+-activated K+ channel complexed with Ca2+/calmodulin. Nature, 410, 1120-1124.
Shannon, K.B. and Li, R. (2000) A myosin light chain mediates the localization of the budding yeast IQGAP-like protein during contractile ring formation. Curr. Biol., 10, 727-730.
Slupsky, C.M., Desautels, M., Huebert, T., Zhao, R., Hemmingsen, S.M. and McIntosh, L.P. (2001) Structure of Cdc4p, a contractile ring protein essential for cytokinesis in Schizosaccharomyces pombe. J. Biol. Chem., 276, 5943-5951.
Stafford, W.F. (1998) Time difference sedimentation velocity analysis of rapidly reversible interacting systems: determination of equilibrium constants by non-linear curve fitting procedures. Biophys. J., 74, A301.
Stafford, W.F., Jacobsen, M.P., Woodhead, J., Craig, R., O'Neall-Hennessey, E. and Szent-Gyorgyi, A.G. (2001) Calcium-dependent structural changes in scallop heavy meromyosin. J. Mol. Biol., 307, 137-147.
Stevens, R.C. and Davis, T.N. (1998) Mlc1p is a light chain for the unconventional myosin Myo2p in Saccharomyces cerevisiae. J. Cell Biol., 142, 711-722.
Terrak, M., Otterbein, L.R., Wu, G., Palecanda, L.A., Lu, R.C. and Dominguez, R. (2002) Crystallization, X-ray characterization and selenomethionine phasing of Mlc1p bound to IQ motifs from myosin V. Acta Crystallogr. D, 58, 1882-1885.
Todd, G.P. and Haschemeyer, R.H. (1981) General solution to the inverse problem of the differential equation of the ultracentrifuge. Proc. Natl Acad. Sci. USA, 78, 6739-6743.
Weeks, C.M. and Miller, R. (1999) The design and implementation of SnB v2.0. J. Appl. Crystallogr., 32, 120-124.
Weissbach,L., Bernards, A. and Herion, D.W. (1998) Binding of myosin essential light chain to the cytoskeleton-associated protein IQGAP1. Biochem. Biophys. Res. Commun., 251, 269-276.
Xie, X., Harrison, D.H., Schlichting, I., Sweet, R.M., Kalabokis, V.N., Szent-Gyorgyi, A.G. and Cohen, C. (1994) Structure of the regulatory domain of scallop myosin at 2.8 Å resolution. Nature, 368, 306-312.
Yin, H., Pruyne, D., Huffaker, T.C. and Bretscher, A. (2000) Myosin V orientates the mitotic spindle in yeast. Nature, 406, 1013-1015.
