MicroRNA, Pm-miR-2305, Participates in Nacre Formation by Targeting Pearlin in Pearl Oyster Pinctada martensii.

Yu Jiao,Rongrong Tian,Ronglian Huang,Zhe Zheng,Xiaodong Du,Qingheng Wang

doi:10.3390/ijms160921442

Yu Jiao, Rongrong Tian + Show 4 more

Open Access

https://doi.org/10.3390/ijms160921442

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Abstract

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.

Highlights

MicroRNAs are short noncoding RNAs that are direct negative regulators of gene expression by binding to specific sequences within a target mRNA [1].MiRNAs repress cellular protein levels to provide a sophisticated parameter of gene regulation that coordinates a broad spectrum of biological processes [2,3]
With the use of the local BLASTN program, the mature sequence of pm-miR-2305 was compared against the transcriptome database of P. martensii [9]
Results showed that only Unigene21313 contained the mature sequence of pm-miR-2305

Summary

Introduction

MicroRNAs (miRNAs) are short (approximately 21-nucleotide) noncoding RNAs that are direct negative regulators of gene expression by binding to specific sequences within a target mRNA [1].MiRNAs repress cellular protein levels to provide a sophisticated parameter of gene regulation that coordinates a broad spectrum of biological processes [2,3]. Inhibition of mRNA translation by miRNAs has proven to be an important regulator of biomineralization, such as bone resorption activity and bone homeostasis, in the adult skeleton [2,3]. MiRNAs control multiple layers of gene regulation for bone development, from the initial response of progenitor cells to the structure and metabolic activity of the mature tissue [2,3]. To investigate the mechanism that controls the biomineralization process in nacre formation, various matrix proteins have been extracted and functionally studied [1,5,6,7]. Nacre formation is a very complex, precise process, and the expression of each related protein is subject to fine regulation [8]

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