Tsiavaliaris, G. and Fujita-Becker, S. and Batra, R. and Levitsky, D.I. and Kull, F.J. and Geeves, M.A. and Manstein, D.J. (2002) Mutations in the relay loop region result in dominant-negative inhibition of myosin II function in Dictyostelium. EMBO reports, 3 (11). pp. 1099-1105. ISSN 1469-221X.
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Dominant-negative inhibition is a powerful genetic tool for the characterization of gene function in vivo, based on the specific impairment of a gene product by the coexpression of a mutant version of the same gene product. We describe the detailed characterization of two myosin constructs containing either point mutations F487A or F506G in the relay region. Dictyostelium cells transformed with F487A or F506G myosin are unable to undergo processes that require myosin II function, including fruiting-body formation, normal cytokinesis and growth in suspension. Our results show that the dominant-negative inhibition of myosin function is caused by disruption of the communication between active site and lever arm, which blocks motor activity completely, and perturbation of the communication between active site and actin-binding site, leading to an approximately 100-fold increase in the mutants' affinity for actin in the presence of ATP.
|Additional information:||1469-221X (Print) Journal Article Research Support, Non-U.S. Gov't|
|Uncontrolled keywords:||Actins/metabolism Animals Calorimetry, Differential Scanning Cell Division/physiology Dictyostelium/cytology/genetics/*physiology Ligands Models, Molecular Molecular Motor Proteins *Mutation Myosin Type II/chemistry/*genetics/*metabolism Protein Conformation Protein Structure, Tertiary|
|Divisions:||Faculties > Science Technology and Medical Studies > School of Biosciences|
|Depositing User:||Michael Geeves|
|Date Deposited:||04 Sep 2008 15:54|
|Last Modified:||14 Jan 2010 14:14|
|Resource URI:||http://kar.kent.ac.uk/id/eprint/4012 (The current URI for this page, for reference purposes)|
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