Ye Chongjun, Jiang Song, Gong Meixia, Min Qin, Fan Manli, Gao Junshan, Meng Yan
School of Life Sciences, Anhui Agricultural University, 130 West Changjiang Road, Hefei 230036, China.
Institute of Sericulture, Anhui Academy of Agricultural Sciences, 15 Huoshan Road, Hefei 230061, China.
Insects. 2020 Aug 12;11(8):523. doi: 10.3390/insects11080523.
The most common type of RNA editing in metazoans is the deamination of adenosine into inosine (A-to-I) catalyzed by the adenosine deaminase acting on the RNA (ADAR) family of proteins. The deletion or dysfunction of ADAR enzymes in higher eukaryotes can affect the efficiency of substrate editing and cause neurological disorders. However, the information concerning A-to-I RNA editing and ADAR members in the silkworm, (BmADAR), is limited. In this study, a first molecular comprehensive cloning and sequence analysis of transcripts was presented. A complete open reading frame (ORF) () was obtained using RT-PCR and RACE and its expression pattern, subcellular localization and A-to-I RNA-editing function on the silkworm () were investigated. Subcellular localization analysis observed that was mainly localized in the nucleus. To further study the A-to-I RNA-editing function of , BmSyt I-pIZ-EGFP was constructed and co-transfected with BmADARa-pIZ-EGFP into BmN cells. The result demonstrates that BmADARa can functionally edit the specific site of Taken together, this study not only provides insight into the function of the first ADAR enzyme in , but also lays foundations for further exploration of the functional domain of BmADARa and its editing substrates and target sites.
后生动物中最常见的RNA编辑类型是由作用于RNA的腺苷脱氨酶（ADAR）家族蛋白催化的腺苷脱氨生成肌苷（A到I）。高等真核生物中ADAR酶的缺失或功能障碍会影响底物编辑效率并导致神经紊乱。然而，关于家蚕中A到I RNA编辑和ADAR成员（BmADAR）的信息有限。在本研究中，首次对转录本进行了分子综合克隆和序列分析。使用RT-PCR和RACE获得了一个完整的开放阅读框（ORF），并研究了其在家蚕中的表达模式、亚细胞定位和A到I RNA编辑功能。亚细胞定位分析表明，其主要定位于细胞核。为了进一步研究其A到I RNA编辑功能，构建了BmSyt I-pIZ-EGFP，并与BmADARa-pIZ-EGFP共转染到BmN细胞中。结果表明，BmADARa可以对进行功能编辑。综上所述，本研究不仅深入了解了家蚕中首个ADAR酶的功能，也为进一步探索BmADARa的功能结构域及其编辑底物和靶位点奠定了基础。
