Sun Xiao-yang, Chen Zi-yan, Hayashi Yoshitaka, Kanou Yasuhiko, Takagishi Yoshiko, Oda Sen-ichi, Murata Yoshiharu
Department of Genetics, Research Institute of Environmental Medicine, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
Gene. 2008 Mar 31;411(1-2):94-102. doi: 10.1016/j.gene.2008.01.013. Epub 2008 Jan 26.
Using forward genetic analysis, we identified the insertion of an intracisternal A particle (IAP) retrotransposon element in the plasma membrane calcium ATPase 2 gene (Pmca2/Atp2b2) in the joggle mouse, a novel mutant that displays ataxic gait by postnatal day 12. Expression of Pmca2 mRNA in the joggle mouse is only 5% of that in the wild type mouse. The insertion is located 15 bp downstream of the donor splice site of the exon containing the initiation codon. Chimeric mRNA composed of the 5'-region of Pmca2 and the IAP element were detected, indicating that some of the primary transcripts are terminated by polyadenylation signals in long terminal repeats of the IAP element. We also identified cryptic splice sites in the IAP element that are likely involved in aberrant splicing of the Pmca2 primary transcripts that leads to rapid degradation of mRNA through nonsense mediated mRNA decay. Ataxia was observed in compound heterozygous mice carrying the joggle mutation and the wriggle mutation, a previously reported missense Pmca2 mutant. Thus, we attributed ataxia in joggle mice to reduced expression of Pmca2, resulting from insertion of the IAP element.
通过正向遗传学分析,我们在“joggle小鼠”的质膜钙ATP酶2基因(Pmca2/Atp2b2)中鉴定出一个脑内池A颗粒(IAP)逆转座子元件的插入,“joggle小鼠”是一种新型突变体,在出生后第12天表现出共济失调步态。“joggle小鼠”中Pmca2 mRNA的表达仅为野生型小鼠的5%。该插入位于包含起始密码子的外显子供体剪接位点下游15 bp处。检测到由Pmca2的5'区域和IAP元件组成的嵌合mRNA,这表明一些初级转录本被IAP元件长末端重复序列中的聚腺苷酸化信号终止。我们还在IAP元件中鉴定出潜在的剪接位点,这些位点可能参与Pmca2初级转录本的异常剪接,导致mRNA通过无义介导的mRNA降解迅速降解。在携带“joggle突变”和“wriggle突变”(先前报道的Pmca2错义突变体)的复合杂合小鼠中观察到共济失调。因此,我们将“joggle小鼠”中的共济失调归因于IAP元件插入导致的Pmca2表达降低。