Transcriptome and proteome analysis of Pinctada margaritifera calcifying mantle and shell: focus on biomineralization.
Transcriptome and proteome analysis of Pinctada margaritifera calcifying mantle and shell: focus on biomineralization.
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DOI：
10.1186/1471-2164-11-613
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发表时间：
2010-11-01
期刊：
影响因子：
4.4
通讯作者：
Montagnani C
中科院分区：
文献类型：
--
作者：
Joubert C;Piquemal D;Marie B;Manchon L;Pierrat F;Zanella-Cléon I;Cochennec-Laureau N;Gueguen Y;Montagnani C
珍珠珠母贝的贝壳是由有机无细胞基质组成的，在生物控制的生物矿化动态过程中起着关键作用。为了增加基因组资源并鉴定与珍珠母贝生物矿化有关的壳基质蛋白，对钙化外套膜进行了高通量表达序列标签（EST）焦磷酸测序，并结合壳的蛋白质组学分析。我们报告了276738个序列的功能分析，导致前所未有的82个珍珠母生物矿化相关的套蛋白序列的目录的构成。发现了当前生物矿化过程的“几丁质-丝素蛋白凝胶-酸性大分子”模型的组分，特别是最近在合浦藻中发现的生物矿化蛋白（Pif-177）的同源物。在这些序列中，我们可以显示定位的其他两个生物矿化蛋白质的转录本，pmarg-aspein和pmarg-pearlin，在两个不同的地区的外地幔上皮，这表明它们的含义在方解石和文石的形成。最后，通过结合EST的方法与蛋白质组学质谱分析的蛋白质分离的P. margaritifera壳有机基质，我们证明了30个序列的存在，包含几乎所有的壳蛋白，已被描述为从壳基质蛋白质分析的珠母贝属。这两种方法的整合，使全球生物矿化组织和钙化结构的组成进行了检查串联的第一次。这项对珍珠贝钙化组织的EST研究是对产珍珠双壳类物种焦磷酸测序的首次描述。我们的研究结果提供了直接的证据，我们的EST数据集涵盖了大部分的多样性的基质蛋白的P. margaritifera壳，但也地幔转录本编码的蛋白质存在于P. margaritifera壳，从而证明他们的暗示在壳的形成。因此，结合转录组学和蛋白质组学方法是鉴定参与生物矿化的蛋白质的有力方法。在这项研究中产生的数据提供了最全面的列表中的生物矿化相关的序列，目前可在原口类物种，并在软体动物生物矿化领域的重大突破。
The shell of the pearl-producing bivalve Pinctada margaritifera is composed of an organic cell-free matrix that plays a key role in the dynamic process of biologically-controlled biomineralization. In order to increase genomic resources and identify shell matrix proteins implicated in biomineralization in P. margaritifera, high-throughput Expressed Sequence Tag (EST) pyrosequencing was undertaken on the calcifying mantle, combined with a proteomic analysis of the shell. We report the functional analysis of 276 738 sequences, leading to the constitution of an unprecedented catalog of 82 P. margaritifera biomineralization-related mantle protein sequences. Components of the current "chitin-silk fibroin gel-acidic macromolecule" model of biomineralization processes were found, in particular a homolog of a biomineralization protein (Pif-177) recently discovered in P. fucata. Among these sequences, we could show the localization of two other biomineralization protein transcripts, pmarg-aspein and pmarg-pearlin, in two distinct areas of the outer mantle epithelium, suggesting their implication in calcite and aragonite formation. Finally, by combining the EST approach with a proteomic mass spectrometry analysis of proteins isolated from the P. margaritifera shell organic matrix, we demonstrated the presence of 30 sequences containing almost all of the shell proteins that have been previously described from shell matrix protein analyses of the Pinctada genus. The integration of these two methods allowed the global composition of biomineralizing tissue and calcified structures to be examined in tandem for the first time. This EST study made on the calcifying tissue of P. margaritifera is the first description of pyrosequencing on a pearl-producing bivalve species. Our results provide direct evidence that our EST data set covers most of the diversity of the matrix protein of P. margaritifera shell, but also that the mantle transcripts encode proteins present in P. margaritifera shell, hence demonstrating their implication in shell formation. Combining transcriptomic and proteomic approaches is therefore a powerful way to identify proteins involved in biomineralization. Data generated in this study supply the most comprehensive list of biomineralization-related sequences presently available among protostomian species, and represent a major breakthrough in the field of molluskan biomineralization.
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影响因子：
3
作者：
Jenny, MJ;Ringwood, AH;Chapman, RW
通讯作者：
Chapman, RW
影响因子：
3
作者：
Jolly, C;Berland, S;Doumenc, D
通讯作者：
Doumenc, D
DOI：
10.1006/bbrc.2000.2274
发表时间：
2000-03-05
影响因子：
3.1
作者：
Kono, M;Hayashi, N;Samata, T
通讯作者：
Samata, T
影响因子：
3.5
作者：
Duplat, D;Puisségur, M;Lopez, E
通讯作者：
Lopez, E
DOI：
10.1046/j.1432-1327.2000.01602.x
发表时间：
2000-08-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Mann, K;Weiss, IM;Fritz, M
通讯作者：
Fritz, M