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Molecular characterization of the Caenorhabditis elegans ALP/Enigma gene alp-1

Molecular characterization of the Caenorhabditis elegans ALP/Enigma gene alp-1. Caroline R. McKeown, Hsiao-Fen Han, and Mary C. Beckerle . Developmental Dynamics Vol 235, Issue 2. Muscle Attachment Sites. -Actinin. Found in dense bodies Crosslinks actin filaments

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Molecular characterization of the Caenorhabditis elegans ALP/Enigma gene alp-1

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  1. Molecular characterization of the Caenorhabditis elegans ALP/Enigma gene alp-1 Caroline R. McKeown, Hsiao-Fen Han, and Mary C. Beckerle Developmental Dynamics Vol 235, Issue 2

  2. Muscle Attachment Sites

  3. -Actinin • Found in dense bodies • Crosslinks actin filaments • Defective -actinin results in cardiomyopathy (not good)

  4. Mammalian ALP • ALP (-actinin associated LIM protein) • Family includes at least 7 proteins • ALP (which has subfamilies including ALP, RIL, CLP36/Elfin, and Mystique) • Enigma • Cypher • All bind either -actinin or -tropomyosin and localize to Z-lines

  5. ALP family domains • LIM (LIN-11, Isl-1, MEC-3) • PDZ (PSD95, Discs-large, ZO-1) • both are known protein binding interfaces

  6. C. elegans ALP is eat-1 • First uncovered in a screen by Leon Avery for mutants with pharyngeal pumping defects • somewhat Unc, also has irregular pharyngeal pumping • SAGE data shows raised expression levels in larval stages • similar expression pattern for -actinin

  7. C. elegans ALP is eat-1

  8. C. elegans ALP is eat-1

  9. ALP-1 Localization • Made GFP translational fusions for alp-1

  10. ALP-1 Localization • All isoforms localize to dense bodies • ALP-1BCD also found at dense plaques • sites of muscle-muscle adhesion at the muscle termini (vinculin co-localization)

  11. PDZ Domain • Sufficient for localization to dense bodies • not sufficient to localize to dense plaques • Made PDZ domain containing (and myc-tagged) construct under influence of heat shock promoter

  12. Conclusions • Not essential for viability • PDZ domain localizes protein to sites of -actinin localization • LIM proteins enable additional protein-protein interactions • many interactions in Y2H interactome project • More study needed for specific role

  13. Layton getting CRAZY!!!

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