Deletions in the extracellular domain of rat trkA lead to an altered differentiative phenotype in neurotrophin responsive cells.
We have investigated the role(s) of conserved sequences in the extracellular domain of rat trkA by generating 5' and 3' deletions and assaying changes in neurotrophin binding, tyrosine kinase activity, and neurite outgrowth. Essential sequences required for both nerve growth factor (NGF) and neurotrophin-3 (NT-3) binding were mapped to the immunoglobulin-like domains. Small deletions in the second immunoglobulin-like domain and in the juxtamembrane region abolished neurotrophin binding. Dose-response curves on cells expressing full-length trkA were identical for NGF and NT-3 (0.1 ng/ml-50 ng/ml) while cells expressing leucine rich motif (LRM) minus receptors required high concentrations of NT-3 (50 ng/ml). Scatchard analysis revealed a loss of high-affinity NT-3, but not NGF, binding to the LRM minus receptor consistent with the neurite dose-response curves. Moreover, cells expressing the LRM minus receptors failed to fasciculate and showed delayed arborization in comparison to cells expressing wild-type trkA, suggesting a possible role for the LRM's in neurotrophin-induced differentiation and in high-affinity NT-3 binding.
MacDonald
J I
JI
Neurodegeneration Group, John P. Robarts Research Institute, London, Ontario, Canada.
Meakin
S O
SO
D000818:Animals; D001667:Binding, Competitive / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D017353:Gene Deletion; D015870:Gene Expression / Q000502:physiology; D008024:Ligands; D016296:Mutagenesis / Q000502:physiology; D009414:Nerve Growth Factors / Q000494:pharmacology*; D020933:Neurotrophin 3; D016716:PC12 Cells / Q000737:chemistry / Q000166:cytology / Q000187:drug effects; D010641:Phenotype; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D051381:Rats; D020917:Receptor, trkA / Q000737:chemistry* / Q000235:genetics*
D008024:Ligands; D009414:Nerve Growth Factors; D020933:Neurotrophin 3; D020917:Receptor, trkA