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

Type Article
Date 2010-11
Language English
Author(s) Joubert Caroline1, Piquemal David2, Marie Benjamin3, Manchon Laurent2, Pierrat Fabien2, Zanella-Cleon Isabelle4, Cochennec-Laureau NathalieORCID5, Gueguen YannickORCID1, Montagnani CarolineORCID1
Affiliation(s) 1 : IFREMER, Lab Biotechnol & Qual Perle, Ctr Oceanol Pacifique, Taravao 98719, Fr Polynesia.
2 : Skuldtech, F-34090 Montpellier, France.
3 : Univ Bourgogne, UMR CNRS Biogeosci 5561, F-21000 Dijon, France.
4 : Univ Lyon 1, IFR BioSci Gerland Lyon Sud 128, UMR CNRS 5086, IBCP, F-69367 Lyon, France.
5 : IFREMER, Ctr Nantes, F-44311 Nantes 03, France.
Source Bmc Genomics (1471-2164) (Biomed Central Ltd), 2010-11 , Vol. 11 , P. 1-13
DOI 10.1186/1471-2164-11-613
WOS© Times Cited 179
Abstract Background: 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. Results: 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. Conclusions: 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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Joubert Caroline, Piquemal David, Marie Benjamin, Manchon Laurent, Pierrat Fabien, Zanella-Cleon Isabelle, Cochennec-Laureau Nathalie, Gueguen Yannick, Montagnani Caroline (2010). Transcriptome and proteome analysis of Pinctada margaritifera calcifying mantle and shell: focus on biomineralization. Bmc Genomics, 11, 1-13. Publisher's official version : , Open Access version :