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

4. 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 Mammalian ALP

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

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

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

10. 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) ALP-1 Localization

11. 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 PDZ Domain

12. 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 Conclusions

13. Layton getting CRAZY!!!