Document Type

Article

Abstract

Kalirin is a multidomain guanine nucleotide exchange factor (GEF) that activates Rho proteins, inducing cytoskeletal rearrangement in neurons. Although much is known about the effects of Kalirin on Rho GTPases and neuronal morphology, little is known about the association of Kalirin with the receptor/signaling systems that affect neuronal morphology. Our experiments demonstrate that Kalirin binds to and colocalizes with the TrkA neurotrophin receptor in neurons. In PC12 cells, inhibition of Kalirin expression using antisense RNA decreased nerve growth factor (NGF)-induced TrkA autophosphorylation and process extension. Kalirin overexpression potentiated neurotrophin-stimulated TrkA autophosphorylation and neurite outgrowth in PC12 cells at a low concentration of NGF. Furthermore, elevated Kalirin expression resulted in catalytic activation of TrkA, as demonstrated by in vitro kinase assays and increased NGF-stimulated cellular activation of Rac, Mek, and CREB. Domain mapping demonstrated that the N-terminal Kalirin pleckstrin homology domain mediates the interaction with TrkA. The effects of Kalirin on TrkA provide a molecular basis for the requirement of Kalirin in process extension from PC12 cells and for previously observed effects on axonal extension and dendritic maintenance. The interaction of TrkA with the pleckstrin homology domain of Kalirin may be one example of a general mechanism whereby receptor/Rho GEF pairings play an important role in receptor tyrosine kinase activation and signal transduction.

Keywords

Animals; Cell differentiation; Cell Differentiation/physiology; Cellular signal transduction; Cyclic AMP Response Element-Binding Protein/metabolism; G proteins; Enzyme activation; G proteins; Genetics; Guanine Nucleotide Exchange Factors/genetics; Guanine Nucleotide Exchange Factors/metabolism; Human genetics; Humans; MAP Kinase Kinase 1/metabolism; Men; Mice; Nerve growth factor; Nerve Growth Factor/genetics; Nerve Growth Factor/metabolism; Neurons/metabolism; PC12 Cells; Protein binding; Protein Isoforms/genetics; Protein Isoforms/metabolism; RNA; RNA; Small Interfering/genetics; RNA; Small Interfering/metabolism; Rats; Receptor; trkA/genetics; Receptor; trkA/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Rho GTPases; Signal Transduction/physiology; Women; rac GTP-Binding Proteins/metabolism

Disciplines

Life Sciences | Molecular Biology | Neurosciences | Structural Biology

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