In the absence of frazzled over-expression of Abelson tyrosine kinase disrupts commissure formation and causes axons to leave the embryonic CNS.
BACKGROUND
In the Drosophila embryonic nerve cord, the formation of commissures require both the chemoattractive Netrin receptor Frazzled (Fra) and the Abelson (Abl) cytoplasmic tyrosine kinase. Abl binds to the cytoplasmic domain of Fra and loss-of-function mutations in abl enhance fra-dependent commissural defects. To further test Abl's role in attractive signaling, we over-expressed Abl in Fra mutants anticipating rescue of commissures.


RESULTS
The Gal4-UAS system was used to pan-neurally over-express Abl in homozygous fra embryos. Surprisingly, this led to a significant decrease in both posterior and anterior commissure formation and induced some commissural and longitudinal axons to project beyond the CNS/PNS border. Re-expressing wild-type Fra, or Fra mutants with a P-motif deleted, revert both commissural and exiting phenotypes, indicating that Fra is required but not a specific P-motif. This is supported by S2 cell experiments demonstrating that Abl binds to Fra independent of any specific P-motif and that Fra continues to be phosphorylated when individual P-motifs are removed. Decreasing midline repulsion by reducing Robo signaling had no effect on the Abl phenotype and the phenotypes still occur in a Netrin mutant. Pan-neural over-expression of activated Rac or Cdc42 in a fra mutant also induced a significant loss in commissures, but axons did not exit the CNS.


CONCLUSIONS
Taken together, these data suggest that Fra activity is required to correctly regulate Abl-dependent cytoskeletal dynamics underlying commissure formation. In the absence of Fra, increased Abl activity appears to be incorrectly utilized downstream of other guidance receptors resulting in a loss of commissures and the abnormal projections of some axons beyond the CNS/PNS border.
Dorsten
Joy N
JN
Department of Biological Sciences, Wayne State University, Detroit, Michigan, United States of America.
Varughese
Bridget E
BE
Karmo
Stephanie
S
Seeger
Mark A
MA
VanBerkum
Mark F A
MF
D020816:Amino Acid Motifs; D000818:Animals; D001369:Axons / Q000378:metabolism*; D002490:Central Nervous System / Q000196:embryology*; D003593:Cytoplasm / Q000378:metabolism; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster; D000939:Epitopes / Q000737:chemistry; D018507:Gene Expression Regulation, Developmental; D009154:Mutation; D000075386:Netrin Receptors; D009474:Neurons / Q000378:metabolism; D010641:Phenotype; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D015398:Signal Transduction
D029721:Drosophila Proteins; D000939:Epitopes; D000075386:Netrin Receptors; D011956:Receptors, Cell Surface; C000617310:fra protein, Drosophila; D011505:Protein-Tyrosine Kinases; C506548:Abl protein, Drosophila