Abstract
Regulator of G-protein signaling (RGS) proteins interact with alpha subunits of heterotrimeric G-proteins via the RGS domain and attenuate their activity by accelerating GTPase activity. RGS2, a member of the RGS family, regulates synaptic development via hereto unknown mechanism. In this study, we found that RGS2 directly interacted with tubulin via a short region at the N-terminus: amino acids 41-60. RGS2 enhanced microtubule polymerization in vitro, and the tubulin binding region was necessary and sufficient for this activity. In Vero cells, polymerization of microtubule was stimulated when peptides containing the tubulin binding region were microinjected. Immunocytochemical analysis showed that endogenous RGS2 was localized at the termini of neurites in differentiated PC12 cells. Over-expression of RGS2 enhanced the nerve growth factor-induced neurite outgrowth in PC12 cells, while specific knock-down of endogenous RGS2 suppressed the neurite outgrowth. These findings demonstrate that RGS2 contributes to the neuronal cell differentiation via regulation of microtubule dynamics.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Binding Sites / genetics
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Brain / metabolism
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COS Cells
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Cell Differentiation / genetics
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Cell Differentiation / physiology
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Chlorocebus aethiops
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Glutathione Transferase / genetics
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Glutathione Transferase / metabolism
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Microscopy, Confocal
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Microscopy, Electron
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Microtubules / metabolism*
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Microtubules / ultrastructure
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Mutation / genetics
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Neurites / metabolism
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Neurites / physiology*
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Neurites / ultrastructure
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PC12 Cells
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Protein Binding
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RGS Proteins / chemistry
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RGS Proteins / genetics
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RGS Proteins / metabolism*
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RNA Interference / physiology
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RNA, Small Interfering / genetics
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Rats
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Transfection / methods
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Tubulin / genetics
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Tubulin / metabolism
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Vero Cells
Substances
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RGS Proteins
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RNA, Small Interfering
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Recombinant Fusion Proteins
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Tubulin
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Green Fluorescent Proteins
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Glutathione Transferase