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PMID:11923418
| Citation |
Yanochko, GM and Yool, AJ (2002) Regulated cationic channel function in Xenopus oocytes expressing Drosophila big brain. J. Neurosci. 22:2530-40 |
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| Abstract |
Big brain (bib) is a neurogenic gene that when mutated causes defects in cell fate determination during Drosophila neurogenesis through an unknown mechanism. The protein Big Brain (BIB) has sequence identity with the major intrinsic protein family that includes the water- and ion-conducting aquaporin channels. We show here that BIB expressed heterologously in Xenopus oocytes provides a voltage-insensitive, nonselective cation channel function with permeability to K+ > Na+ > tetraethylammonium. The conductance, activated in response to endogenous signaling pathways in BIB-expressing oocytes, is decreased after treatment with 20 microm insulin and is enhanced with 10 microm lavendustin A, a tyrosine kinase inhibitor. Western blot analysis confirms that BIB is tyrosine-phosphorylated. Both tyrosine phosphorylation and the potentiating effect of lavendustin A are removed by partial deletion of the C terminus (amino acids 317-700). Current activation is not observed in control oocytes or in oocytes expressing a nonfunctional mutant (BIB E71N) that appears to be expressed on the plasma membrane by confocal microscopy and Western blotting. These results indicate that BIB can participate in tyrosine kinase-regulated transmembrane signaling and may suggest a role for membrane depolarization in the neurogenic function of BIB in early development. |
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| Keywords |
Animals; Cations/metabolism; Cell Membrane Permeability/drug effects; Cell Membrane Permeability/physiology; Dose-Response Relationship, Drug; Drosophila; Drosophila Proteins; Enzyme Inhibitors/pharmacology; Gene Expression; Insulin/pharmacology; Ion Channels/drug effects; Ion Channels/metabolism; Ion Transport/drug effects; Ion Transport/physiology; Membrane Potentials/drug effects; Membrane Potentials/physiology; Membrane Proteins/genetics; Membrane Proteins/metabolism; Microinjections; Mutagenesis, Site-Directed; Oocytes/cytology; Oocytes/drug effects; Oocytes/metabolism; Osmolar Concentration; Patch-Clamp Techniques; Phosphorylation; RNA, Complementary/administration & dosage; Signal Transduction/physiology; Structure-Activity Relationship; Transfection; Water/metabolism; Xenopus |
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Significance
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| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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