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PU14, a Novel Matrix Protein, Participates in Pearl Oyster, Pinctada Fucata , Shell Formation
Marine Biotechnology ( IF 3 ) Pub Date : 2021-03-10 , DOI: 10.1007/s10126-020-10014-3
Yinghui Ji 1 , Xue Yang 1 , Dong Yang 1 , Rongqing Zhang 1, 2, 3
Affiliation  

Biomineralization is a widespread biological process, involved in the formation of shells, teeth, and bones. Shell matrix proteins have been widely studied for their importance during shell formation. In 2015, our group identified 72 unique shell matrix proteins in Pinctada fucata, among which PU14 is a matrix protein detected in the soluble fraction that solely exists in the prismatic layer. However, the function of PU14 is still unclear. In this study, the full-length cDNA sequence of PU14 was obtained and functional analyses of PU14 protein during shell formation were performed. The deduced protein has a molecular mass of 77.8 kDa and an isoelectric point of 11.34. The primary protein structure contains Gln-rich and random repeat units, which are typical characteristics of matrix protein and indicate its potential function during shell formation. In vivo and in vitro experiments indicated PU14 has prismatic layer functions during shell formation. The tissue expression patterns showed that PU14 was mainly expressed in the mantle tissue, which is consistent with prismatic layer formation. Notching experiments suggested that PU14 responded to repair and regenerate the injured shell. After inhibiting gene expression by injecting PU14-specific double-stranded RNA, the inner surface of the prismatic layer changed significantly and became rougher. Further, in vitro experiments showed that recombinant protein rPU14 impacted calcite crystal morphology. Taken together, characterization and functional analyses of a novel matrix protein, PU14, provide new insights about basic matrix proteins and their functions during shell formation.



中文翻译:

PU14,一种新型基质蛋白,参与珍珠牡蛎、珠母贝、贝壳形成

生物矿化是一种广泛的生物过程,涉及贝壳、牙齿和骨骼的形成。壳基质蛋白因其在壳形成过程中的重要性而被广泛研究。2015年,我们课题组在紫珠母贝中鉴定出72种独特的壳基质蛋白,其中PU14是一种基质蛋白,在可溶性部分中检测到,仅存在于棱柱层中。然而,PU14的功能仍不清楚。在这项研究中,获得了 PU14 的全长 cDNA 序列,并进行了壳形成过程中 PU14 蛋白的功能分析。推导出的蛋白质的分子量为 77.8 kDa,等电点为 11.34。蛋白质一级结构包含富含 Gln 和随机重复单元,这是基质蛋白的典型特征,表明其在壳形成过程中的潜在功能。体内外实验表明 PU14 在壳形成过程中具有棱柱层功能。组织表达模式表明PU14主要在地幔组织中表达,这与棱柱形层的形成一致。缺口实验表明PU14对修复和再生受伤的外壳做出了反应。通过注射PU14特异性双链RNA抑制基因表达后,棱柱层的内表面发生显着变化并变得更加粗糙。此外,体外实验表明重组蛋白 rPU14 影响方解石晶体形态。总之,新型基质蛋白 PU14 的表征和功能分析为基本基质蛋白及其在壳形成过程中的功能提供了新的见解。

更新日期:2021-03-10
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