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Functional characterization and molecular mechanism exploration of three granulin epithelin precursor splice variants in biomineralization of the pearl oyster Pinctada fucata
Zhao, Mi; He, Maoxian; Huang, Xiande; Wang, Qi; Shi, Yu; Zhao, M (reprint author), Chinese Acad Sci, South China Sea Inst Oceanol, CAS Key Lab Trop Marine Bioresources & Ecol, Guangzhou 510301, Guangdong, Peoples R China.
2016
Source PublicationMOLECULAR GENETICS AND GENOMICS
Volume291Issue:1Pages:399-409
AbstractThe granulin/epithelin precursor (GEP) encodes a glycoprotein precursor which exhibits pleiotropic tissue growth factor activity with multiple functions. Here, GEP was isolated and its role in the shell biomineralization process of the pearl oyster Pinctada fucata was investigated. Three forms of GEP mRNA were isolated from the pearl oyster (designated PfGEP-1, PfGEP-2 and PfGEP-3). Genomic DNA flanking the splicing region of the PfGEP variants was sequenced and it was found that PfGEP-2 splices out Exon 4, whereas PfGEP-3 splices out Exon 3 compared to PfGEP-1. PfGEP-1 (1505 amino acids) consists of 18 granulin domains, whereas PfGEP-2 (1459 amino acids) and PfGEP-3 (1471 amino acids) consist of 17.5 granulin domains, respectively. Analyses of PfGEP-1 and PfGEP-3 mRNA showed differential patterns in the tissues and developmental stages. Western blotting results showed that the three splice variants can translate to proteins in HEK293T cells. A knockdown experiment using PfGEP dsRNA showed decreased PfGEP-1/PfGEP-3 and PfMSX mRNA, and irregular crystallization of the nacreous layer using scanning electron microscopy. In luciferase assays, co-transfection of PfGEP-1 could activate as well as repress luciferase expression of the reporter plasmid driven by the PfMSX promoter, whereas PfGEP-3 stimulated the expression, elucidating the molecular mechanisms involved in the correlation between PfGEP and PfMSX. These results suggested that GEP variants might function differently during the biomineralization process, which provides new knowledge on the mechanism regulating nacre formation.
Department[Zhao, Mi; He, Maoxian; Huang, Xiande; Wang, Qi; Shi, Yu] Chinese Acad Sci, South China Sea Inst Oceanol, CAS Key Lab Trop Marine Bioresources & Ecol, Guangzhou 510301, Guangdong, Peoples R China; [Zhao, Mi; Wang, Qi] Univ Chinese Acad Sci, Beijing 100049, Peoples R China ; LMB
KeywordGep Splice Variants Biomineralization Gep-msx Signaling Pathway Pinctada Fucata
Subject AreaBiochemistry & Molecular Biology ; Genetics & Heredity
Document Type期刊论文
Identifierhttp://ir.scsio.ac.cn/handle/344004/15504
Collection中科院海洋生物资源可持续利用重点实验室
Corresponding AuthorZhao, M (reprint author), Chinese Acad Sci, South China Sea Inst Oceanol, CAS Key Lab Trop Marine Bioresources & Ecol, Guangzhou 510301, Guangdong, Peoples R China.
Recommended Citation
GB/T 7714
Zhao, Mi,He, Maoxian,Huang, Xiande,et al. Functional characterization and molecular mechanism exploration of three granulin epithelin precursor splice variants in biomineralization of the pearl oyster Pinctada fucata[J]. MOLECULAR GENETICS AND GENOMICS,2016,291(1):399-409.
APA Zhao, Mi,He, Maoxian,Huang, Xiande,Wang, Qi,Shi, Yu,&Zhao, M .(2016).Functional characterization and molecular mechanism exploration of three granulin epithelin precursor splice variants in biomineralization of the pearl oyster Pinctada fucata.MOLECULAR GENETICS AND GENOMICS,291(1),399-409.
MLA Zhao, Mi,et al."Functional characterization and molecular mechanism exploration of three granulin epithelin precursor splice variants in biomineralization of the pearl oyster Pinctada fucata".MOLECULAR GENETICS AND GENOMICS 291.1(2016):399-409.
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