title: Structural basis for the interaction of the myosin light chain Mlc1p with the myosin V Myo2p IQ motifs.
abstract: Calmodulin, regulatory, and essential myosin light chain are evolutionary conserved proteins that, by binding to IQ motifs of target proteins, regulate essential intracellular processes among which are efficiency of secretory vesicles release at synapsis, intracellular signaling, and regulation of cell division. The yeast Saccharomyces cerevisiae calmodulin Cmd1 and the essential myosin light chain Mlc1p share the ability to interact with the class V myosin Myo2p and Myo4 and the class II myosin Myo1p. These myosins are required for vesicle, organelle, and mRNA transport, spindle orientation, and cytokinesis. We have used the budding yeast model system to study how calmodulin and essential myosin light chain selectively regulate class V myosin function. NMR structural analysis of uncomplexed Mlc1p and interaction studies with the first three IQ motifs of Myo2p show that the structural similarities between Mlc1p and the other members of the EF-hand superfamily of calmodulin-like proteins are mainly restricted to the C-lobe of these proteins. The N-lobe of Mlc1p presents a significantly compact and stable structure that is maintained both in the free and complexed states. The Mlc1p N-lobe interacts with the IQ motif in a manner that is regulated both by the IQ motifs sequence as well as by light chain structural features. These characteristic allows a distinctive interaction of Mlc1p with the first IQ motif of Myo2p when compared with calmodulin. This finding gives us a novel view of how calmodulin and essential light chain, through a differential binding to IQ1 of class V myosin motor, regulate this activity during vegetative growth and cytokinesis.
authors.affiliation: Department of Chemical Science and Technology, Università di Roma Tor Vergata, Via della Ricerca Scientifica 1, 00133 Rome, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Pennestri
authors.forename: Sonia
authors.initials: S
authors.lastname: Melino
authors.forename: Gian Marco
authors.initials: GM
authors.lastname: Contessa
authors.forename: Elena Caroli
authors.initials: EC
authors.lastname: Casavola
authors.forename: Maurizio
authors.initials: M
authors.lastname: Paci
authors.forename: Antonella
authors.initials: A
authors.lastname: Ragnini-Wilson
authors.forename: Daniel O
authors.initials: DO
authors.lastname: Cicero
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002147:Calmodulin / Q000737:chemistry; D002455:Cell Division; D003001:Cloning, Molecular; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018995:Myosin Heavy Chains / Q000737:chemistry*; D018994:Myosin Light Chains / Q000737:chemistry* / Q000378:metabolism; D024701:Myosin Type V / Q000737:chemistry*; D009218:Myosins / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002147:Calmodulin; C113979:MLC1 protein, S cerevisiae; C077512:MYO2 protein, S cerevisiae; D018994:Myosin Light Chains; D029701:Saccharomyces cerevisiae Proteins; D024701:Myosin Type V; D018995:Myosin Heavy Chains; D009218:Myosins