Functional Complementation of Glra1spd-ot, a Glycine Receptor Subunit Mutant, by Independently Expressed C-Terminal Domains
Functional Complementation of Glra1spd-ot, a Glycine Receptor Subunit Mutant, by Independently Expressed C-Terminal Domains
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通过独立表达的 C 端结构域对甘氨酸受体亚基突变体 Glra1spd-ot 进行功能互补
DOI：
10.1523/jneurosci.4400-08.2009
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
C. Becker
中科院分区：
文献类型：
--
作者：
C. Villmann;Jana Oertel;Zhan;M. Hollmann;R. Sprengel;Kristina Becker;H. Breitinger;C. Becker
振荡小鼠（Glra 1 spd-ot）在甘氨酸受体α1亚基基因中携带9 bp微缺失和2 bp微插入，导致CNS中功能性α1多肽的缺失和出生后3周的致死性。根据Glra 1基因外显子9中两个剪接受体位点的差异使用，突变等位基因编码截短的α1亚基（spdot-trc）或具有C末端错义序列的多肽（spdot-elg）。在重组表达过程中，两种剪接变体都不能形成离子通道。在互补研究中，编码缺失的C-末端序列的尾部构建体与两种突变体共表达。与spdot-trc共表达产生甘氨酸门控离子通道。通过包含野生型基序RRKRRH来增加拯救效率。在培养的脊髓神经元振子纯合子，病毒感染重组C-末端尾结构导致内源性α1抗原的外观。α1突变体的C-末端尾部多肽的功能拯救主张Cys-环受体家族成员的模块化亚基结构。
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 the Glra1 gene, 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.
影响因子：
4.8
作者：
Zolotukhin, S;Potter, M;Snyder, RO
通讯作者：
Snyder, RO