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Cited by 6Protein Kinase Regulation and GTPase SignalingCellular Mechanics and InteractionsCell Adhesion Molecules Research

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The dual Ras-association domains of Drosophila Canoe have differential roles in linking cell junctions to the cytoskeleton during morphogenesis

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McParland, Emily D., Gurley, Noah J., Wolfsberg, Leah R., Butcher, T. Amber, Bhattarai, Abhi, Jensen, Corbin C., Johnson, Ruth I., Slep, Kevin C., Peifer, Mark·2024·Journal of Cell Science
DOI 10.1242/jcs.263546

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Abstract

During development cells must change shape and move without disrupting dynamic tissue architecture. This requires robust linkage of cell–cell adherens junctions to the force-generating actomyosin cytoskeleton. Drosophila Canoe and mammalian afadin play key roles in the regulation of such linkages. One central task for the field is defining mechanisms by which upstream inputs from Ras-family GTPases regulate Canoe and afadin. These proteins are unusual in sharing two tandem Ras-association (RA) domains – RA1 and RA2 – which when deleted virtually eliminate Canoe function. Work in vitro has suggested that RA1 and RA2 differ in GTPase affinity, but their individual functions in vivo remain unknown. Combining bioinformatic and biochemical approaches, we find that both RA1 and RA2 bind to active Rap1 with similar affinities, and that their conserved N-terminal extensions enhance binding. We created Drosophila canoe mutants to test RA1 and RA2 function in vivo. Despite their similar affinities for Rap1, RA1 and RA2 play strikingly different roles. Deleting RA1 virtually eliminates Canoe function, whereas mutants lacking RA2 are viable and fertile but have defects in junctional reinforcement in embryos and during pupal eye development. These data significantly expand our understanding of the regulation of adherens junction–cytoskeletal linkages.

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Metadata

Authors
McParland, E. D., Gurley, N. J., Wolfsberg, L. R., Butcher, T. A., Bhattarai, A., Jensen, C. C., Johnson, R. I., Slep, K. C., & Peifer, M.
Affiliations
  • University of North Carolina at Chapel Hill (US)
  • Wesleyan University (US)
Corresponding author
  • Slep, Kevin C.
  • Peifer, Mark
ORCID
Venue
Journal of Cell Science
Publisher
The Company of Biologists
Published
2024-12-01
Volume
137
Issue
23
ISSN
  • 0021-9533 (ISSN-L)
  • 1477-9137
DOI
10.1242/jcs.263546
OpenAlex
W4403768074
Cited by
6
References
1
Citations by year
2026: 2 · 2025: 3 · 2024: 1
FWCI
0.81
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Cell Science (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
Funders
  • National Institutes of Health (R35 GM118096, R15 GM114729)
  • University of North Carolina at Chapel Hill
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
Keywords
canoes, afadin, Ras association domain, Rap1, adherens junctions, actomyosin cytoskeleton, cell shape changes, cell migration, Drosophila embryo
MeSH terms
Protein Domains, Animals, Cytoskeleton, Drosophila melanogaster, Intercellular Junctions, Morphogenesis, Protein Binding, ras Proteins, Adherens Junctions, Drosophila Proteins

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