Mechanism of disease caused by deficiency of degradation of tRNA intron in mitochondria

编码ACR-毒素受体的ACRS基因在毒素不敏感的柑橘品种线粒体中降解。为了鉴定ACRSmRNA降解蛋白与ACRS mRNA结合蛋白AmBP30结合蛋白复合体的组成蛋白，以AmBP30为诱饵，用AmBP30和Y2H选择蛋白抗体进行免疫沉淀，通过飞行时间-质谱仪对沉淀蛋白进行分析，确定多个蛋白质是ACRSmRNA降解蛋白复合体的组成蛋白。此外，通过靶基因干扰和RNA沉默，证实了多个基因在ACR和ACT-毒素生物合成中的作用。

ACRS gene encoding ACR-toxin receptor is degraded in mitochondria from the toxin-insensitive citrus cultivars. To identify the component proteins consisting ACRSmRNA degradation protein complex with AmBP30, which is ACRS mRNA-binding protein, yeast two hybrid with AmBP30 as the bait and immuno-precipitation using AmBP30 and Y2H-selected protein antibodies following the precipitated protein analysis by TOF-MS confirmed multiple proteins as the components of ACRSmRNA degradation protein complex. Furthermore, roles of multiple genes in ACR and ACT-toxins biosynthesis were confirmed using target-gene disruption and RNA silencing.