Transcriptome and proteome analysis of Pinctada margaritifera calcifying mantle and shell: focus on biomineralization

BMC Genomics

Research Article

Isabelle Zanella-Cléon¹  Yannick Gueguen²  Caroline Joubert²  Caroline Montagnani²  Nathalie Cochennec-Laureau³  David Piquemal⁴  Fabien Pierrat⁴  Laurent Manchon⁴  Benjamin Marie⁵ 

[1] IFR 128 BioSciences Gerland-Lyon Sud; UMR 5086 CNRS; IBCP, Institut de Biologie et Chimie des Protéines, Université de Lyon 1, 7 passage du Vercors, F-69367, Lyon, France;Ifremer - Laboratoire de Biotechnologie et de Qualité de la Perle, Centre Océanologique du Pacifique - BP 7004, 98719, Taravao, Polynesia, French;Ifremer, Centre de Nantes- rue de l'Ile d'Yeu - BP 21105, 44311, Nantes cedex 03, France;Skuldtech, 134, rue du Curat - Bat. Amarante, 34090, Montpellier, France;UMR CNRS 5561 Biogéosciences, Université de Bourgogne, 6, bd Gabriel, 21000, Dijon, France;

关键词: Silk Fibroin;    Pearl Oyster;    Whey Acidic Protein;    Nacreous Layer;    Biomineralization Process;   

DOI  :  10.1186/1471-2164-11-613

 received in 2010-07-08, accepted in 2010-11-01,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundThe 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.ResultsWe 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.ConclusionsThis 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.

【 授权许可】

Unknown   
© Joubert et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.