Functional complementation of Glra1(spd-ot), a glycine receptor subunit mutant, by independently expressed C-terminal domains
| Autor: | Rolf Sprengel, Hans-Georg Breitinger, Zhan-Lu Ma-Högemeier, Cord-Michael Becker, Jana Oertel, Kristina Becker, Michael Hollmann, Carmen Villmann |
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| Rok vydání: | 2009 |
| Předmět: |
Models
Molecular Patch-Clamp Techniques Protein subunit Mutant Mice Transgenic Biology Transfection Interleukin 10 receptor alpha subunit law.invention Membrane Potentials Exon Mice Receptors Glycine law Chlorocebus aethiops Animals Humans Biotinylation Gene Glycine receptor Cells Cultured Neurons General Neuroscience Articles Embryo Mammalian Molecular biology Protein Structure Tertiary Complementation Luminescent Proteins Spinal Cord Mutation Recombinant DNA Mutagenesis Site-Directed Ion Channel Gating |
| Zdroj: | The Journal of Neuroscience : the Official Journal of the Society for Neuroscience |
| ISSN: | 1529-2401 |
| Popis: | The oscillator mouse (Glra1spd-ot) carries a 9 bp microdeletion plus a 2 bp microinsertion in the glycine receptor α1 subunit gene, resulting in the absence of functional α1 polypeptides from the CNS and lethality 3 weeks after birth. Depending on differential use of two splice acceptor sites in exon 9 of theGlra1gene, the mutant allele encodes either a truncated α1 subunit (spdot-trc) or a polypeptide with a C-terminal missense sequence (spdot-elg). During recombinant expression, both splice variants fail to form ion channels. In complementation studies, a tail construct, encoding the deleted C-terminal sequence, was coexpressed with both mutants. Coexpression with spdot-trc produced glycine-gated ion channels. Rescue efficiency was increased by inclusion of the wild-type motif RRKRRH. In cultured spinal cord neurons from oscillator homozygotes, viral infection with recombinant C-terminal tail constructs resulted in appearance of endogenous α1 antigen. The functional rescue of α1 mutants by the C-terminal tail polypeptides argues for a modular subunit architecture of members of the Cys-loop receptor family. |
| Databáze: | OpenAIRE |
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