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The transcription factor Pf- POU3F4 regulates expression of the matrix protein genes Aspein and Prismalin-14 in pearl oyster ( Pinctada fucata).
- Source :
-
FEBS Journal . May2016, Vol. 283 Issue 10, p1962-1978. 17p. - Publication Year :
- 2016
-
Abstract
- Matrix proteins play key roles in shell formation in the pearl oyster, but little is known about how these proteins are regulated. Here, two POU domain family members, Pf- POU2F1 and Pf- POU3F4, were cloned and characterized. Functional domain analysis revealed that both them have conserved POUS and POUH domains; these domains are important for transcription factor function. The tissue distributions of Pf- POU2F1 and Pf- POU3F4 mRNAs in pearl oyster revealed different expression patterns, and the expression of Pf- POU3F4 m RNA was relatively high in the mantle. The promoters of the matrix protein genes Aspein and Prismalin-14 were cloned using genome-walking PCR. Relatively high transcriptional activities of these promoters were detected in HEK-293T cells. In transient co-transfection assays, Pf- POU3F4 greatly up-regulated the promoter activities of the Aspein and Prismalin-14 genes in a dose-dependent manner. Structural integrity of Pf- POU3F4 was essential for its activation function. One region of the Aspein gene promoter, −181 to −77 bp, and two binding sites in the Prismalin-14 gene promoter, −359 to −337 bp and −100 to −73 bp, were required for activation of Pf- POU3F4. An electrophoresis mobility shift assay demonstrated that Pf- POU3F4 directly bound these sites. Pf- POU3F4 knockdown led to a decrease in Aspein and Prismalin-14 gene expression. Furthermore, expression levels for the Pf- POU3F4 gene were similar to those of the Aspein and Prismalin-14 genes during five development stages. Taken together, these results suggest that the transcription factor Pf- POU3F4 regulates expression of the matrix protein genes Aspein and Prismalin-14 in pearl oyster. Database The nucleotide sequence data of Pf-POU2F1 is available in the GenBank databases under the accession number . The nucleotide sequence data of Pf-POU3F4 is available in the GenBank databases under the accession number . The nucleotide sequence data of Aspein gene promoter is available in the GenBank databases under the accession number . The nucleotide sequence data of Prismalin-14 gene promoter is available in the GenBank databases under the accession number . [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 1742464X
- Volume :
- 283
- Issue :
- 10
- Database :
- Academic Search Index
- Journal :
- FEBS Journal
- Publication Type :
- Academic Journal
- Accession number :
- 115595753
- Full Text :
- https://doi.org/10.1111/febs.13716