MicroRNA, Pm-miR-2305, Participates in Nacre Formation by Targeting Pearlin in Pearl Oyster Pinctada martensii.
MicroRNA, Pm-miR-2305, Participates in Nacre Formation by Targeting Pearlin in Pearl Oyster Pinctada martensii.
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MicroRNA，Pm-miR-2305，通过靶向马氏珠母贝中的珍珠蛋白参与珍珠层形成
DOI：
10.3390/ijms160921442
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发表时间：
2015-09-07
影响因子：
5.6
通讯作者：
Huang R
中科院分区：
文献类型：
--
作者：
Jiao Y;Zheng Z;Tian R;Du X;Wang Q;Huang R
microRNA（miRNAs）是一类非编码RNA分子，对靶基因起负调控作用。在我们之前的研究中，通过Solexa深度测序技术在马氏珠母贝中鉴定出258个pm-miRNAs。Pm-miR-2305是已鉴定的在生物矿化中具有潜在功能的pm-miRNAs之一。在本研究中，预测pm-miR-2305的前体为96 bp，含有特征性的发夹结构。Stem-loop qRT-PCR分析表明，pm-miR-2305在马氏珠母贝的所有组织（闭壳肌、鳃、外套膜、肝胰腺、足和性腺）中组成型表达，并且在足中高度表达。通过模拟注射pm-miR-2305到马氏珠母贝的肌肉中，在外套膜中过量表达pm-miR-2305后，珍珠层表现出无序生长，如通过扫描电子显微镜检测到的。双荧光素酶报告基因检测结果表明，pm-miR-2305模拟物能显著抑制含有pearlin基因3′UTR的报告基因的荧光素酶活性。Western blot分析表明，pm-miR-2305过表达后，外套膜组织中pearlin蛋白表达下调。因此，我们的数据表明，pm-miR-2305通过靶向珍珠质参与了珍珠层的形成。
MicroRNAs (miRNAs) are noncoding RNA molecules that function as negative regulators of target genes. In our previous research, 258 pm-miRNAs were identified in Pinctada martensii by Solexa deep sequencing. Pm-miR-2305 was one of the identified pm-miRNAs with a potential function in biomineralization. In the present study, the precursor of pm-miR-2305 was predicted with 96 bp, containing a characteristic hairpin structure. Stem-loop qRT-PCR analysis indicated that pm-miR-2305 was constitutively expressed in all the tissues (adductor muscle, gill, mantle, hepatopancreas, foot, and gonad) of P. martensii and was highly expressed in the foot. After the over-expression of pm-miR-2305 in the mantle by mimics injection into the muscle of P. martensii, nacre demonstrated disorderly growth, as detected by scanning electron microscopy. Dual luciferase reporter gene assay indicated that pm-miR-2305 mimics could significantly inhibit the luciferase activity of the reporter containing the 3′UTR of the pearlin gene. Western blot analysis demonstrated that the protein expression of pearlin was down-regulated in the mantle tissue after the over-expression of pm-miR-2305. Therefore, our data showed that pm-miR-2305 participated in nacre formation by targeting pearlin in P. martensii.