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Conformational changes in twitchin kinase in vivo revealed by FRET imaging of freely moving C. elegans

Conformational changes in twitchin kinase in vivo revealed by FRET imaging of freely moving C. elegans

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PORTO, Daniel, Yohei MATSUNAGA, Barbara FRANKE, Rhys M. WILLIAMS, Hiroshi QADOTA, Olga MAYANS, Guy M. BENIAN, Hang LU, 2021. Conformational changes in twitchin kinase in vivo revealed by FRET imaging of freely moving C. elegans. In: eLife. eLife Sciences Publications. 10, e66862. eISSN 2050-084X. Available under: doi: 10.7554/eLife.66862

@article{Porto2021Confo-55521, title={Conformational changes in twitchin kinase in vivo revealed by FRET imaging of freely moving C. elegans}, year={2021}, doi={10.7554/eLife.66862}, volume={10}, journal={eLife}, author={Porto, Daniel and Matsunaga, Yohei and Franke, Barbara and Williams, Rhys M. and Qadota, Hiroshi and Mayans, Olga and Benian, Guy M. and Lu, Hang}, note={Article Number: e66862} }

<rdf:RDF xmlns:dcterms="http://purl.org/dc/terms/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#" xmlns:bibo="http://purl.org/ontology/bibo/" xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#" xmlns:foaf="http://xmlns.com/foaf/0.1/" xmlns:void="http://rdfs.org/ns/void#" xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > <rdf:Description rdf:about="https://kops.uni-konstanz.de/rdf/resource/123456789/55521"> <dcterms:title>Conformational changes in twitchin kinase in vivo revealed by FRET imaging of freely moving C. elegans</dcterms:title> <dc:creator>Matsunaga, Yohei</dc:creator> <dc:contributor>Matsunaga, Yohei</dc:contributor> <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/55521/1/Porto_2-iks77b77s1kc4.pdf"/> <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by/4.0/"/> <dc:contributor>Williams, Rhys M.</dc:contributor> <dc:contributor>Lu, Hang</dc:contributor> <dc:creator>Benian, Guy M.</dc:creator> <dc:contributor>Franke, Barbara</dc:contributor> <dc:language>eng</dc:language> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2021-11-12T09:38:39Z</dcterms:available> <dc:creator>Qadota, Hiroshi</dc:creator> <dc:contributor>Benian, Guy M.</dc:contributor> <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/> <foaf:homepage rdf:resource="http://localhost:8080/jspui"/> <dc:creator>Mayans, Olga</dc:creator> <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dc:contributor>Mayans, Olga</dc:contributor> <dcterms:abstract xml:lang="eng">The force-induced unfolding and refolding of proteins is speculated to be a key mechanism in the sensing and transduction of mechanical signals in the living cell. Yet, little evidence has been gathered for its existence in vivo. Prominently, stretch-induced unfolding is postulated to be the activation mechanism of the twitchin/titin family of autoinhibited sarcomeric kinases linked to the mechanical stress response of muscle. To test the occurrence of mechanical kinase activation in living working muscle, we generated transgenic Caenorhabditis elegans expressing twitchin containing FRET moieties flanking the kinase domain and developed a quantitative technique for extracting FRET signals in freely moving C. elegans, using tracking and simultaneous imaging of animals in three channels (donor fluorescence, acceptor fluorescence, and transmitted light). Computer vision algorithms were used to extract fluorescence signals and muscle contraction states in each frame, in order to obtain fluorescence and body curvature measurements with spatial and temporal precision in vivo. The data revealed statistically significant periodic changes in FRET signals during muscle activity, consistent with a periodic change in the conformation of twitchin kinase. We conclude that stretch-unfolding of twitchin kinase occurs in the active muscle, whereby mechanical activity titrates the signaling pathway of this cytoskeletal kinase. We anticipate that the methods we have developed here could be applied to obtaining in vivo evidence for force-induced conformational changes or elastic behavior of other proteins not only in C. elegans but in other animals in which there is optical transparency (e.g., zebrafish).</dcterms:abstract> <dc:rights>Attribution 4.0 International</dc:rights> <dcterms:issued>2021</dcterms:issued> <dc:creator>Porto, Daniel</dc:creator> <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/55521"/> <dc:contributor>Qadota, Hiroshi</dc:contributor> <dc:contributor>Porto, Daniel</dc:contributor> <dc:creator>Williams, Rhys M.</dc:creator> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2021-11-12T09:38:39Z</dc:date> <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/rdf/resource/123456789/28"/> <dc:creator>Franke, Barbara</dc:creator> <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/55521/1/Porto_2-iks77b77s1kc4.pdf"/> <dc:creator>Lu, Hang</dc:creator> </rdf:Description> </rdf:RDF>

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