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• W2069013215 abstract "SGF-2 binds to promoter elements governing posterior silk gland-specific expression of the fibroin gene in Bombyx mori. We purified SGF-2 and showed that SGF-2 contains at least four gene products: the silkworm orthologues of LIM homeodomain protein Awh, LIM domain-binding protein (Ldb), a sequence-specific single-stranded DNA-binding protein (Lcaf), and the silk protein P25/fibrohexamerin (fhx). Using co-expression of these factors in Sf9 cells, Awh, Ldb, and Lcaf proteins were co-purified as a ternary complex that bound to the enhancer sequence in vitro. Lcaf interacts with Ldb as well as Awh through the conserved regions to mediate transcriptional activation in yeast. Misexpression of Awh in transgenic silkworms induces ectopic expression of the fibroin gene in the middle silk glands, where Ldb and Lcaf are expressed. Taken together, this study demonstrates that SGF-2 is a multisubunit activator complex containing Awh. Moreover, our results suggest that the Ldb·Lcaf protein complex serves as a scaffold to facilitate communication between transcriptional control elements.Background: Silk gland factor-2 (SGF-2) is a key factor regulating tissue-specific expression of the fibroin gene.Results: SGF-2 is a 1.1-MDa heteromeric complex containing Awh, Ldb, Lcaf, and fibrohexamerin proteins.Conclusion: Awh, Ldb, and Lcaf interact functionally in SGF-2 to control fibroin gene expression.Significance: This study provides new insight into the functional role of single-stranded DNA-binding proteins in protein-protein interaction and transcriptional regulation. SGF-2 binds to promoter elements governing posterior silk gland-specific expression of the fibroin gene in Bombyx mori. We purified SGF-2 and showed that SGF-2 contains at least four gene products: the silkworm orthologues of LIM homeodomain protein Awh, LIM domain-binding protein (Ldb), a sequence-specific single-stranded DNA-binding protein (Lcaf), and the silk protein P25/fibrohexamerin (fhx). Using co-expression of these factors in Sf9 cells, Awh, Ldb, and Lcaf proteins were co-purified as a ternary complex that bound to the enhancer sequence in vitro. Lcaf interacts with Ldb as well as Awh through the conserved regions to mediate transcriptional activation in yeast. Misexpression of Awh in transgenic silkworms induces ectopic expression of the fibroin gene in the middle silk glands, where Ldb and Lcaf are expressed. Taken together, this study demonstrates that SGF-2 is a multisubunit activator complex containing Awh. Moreover, our results suggest that the Ldb·Lcaf protein complex serves as a scaffold to facilitate communication between transcriptional control elements. Background: Silk gland factor-2 (SGF-2) is a key factor regulating tissue-specific expression of the fibroin gene. Results: SGF-2 is a 1.1-MDa heteromeric complex containing Awh, Ldb, Lcaf, and fibrohexamerin proteins. Conclusion: Awh, Ldb, and Lcaf interact functionally in SGF-2 to control fibroin gene expression. Significance: This study provides new insight into the functional role of single-stranded DNA-binding proteins in protein-protein interaction and transcriptional regulation." @default.
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• W2069013215 date "2013-11-01" @default.
• W2069013215 modified "2023-10-12" @default.
• W2069013215 title "Silk Gland Factor-2, Involved in Fibroin Gene Transcription, Consists of LIM Homeodomain, LIM-interacting, and Single-stranded DNA-binding Proteins" @default.
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• W2069013215 doi "https://doi.org/10.1074/jbc.m113.514471" @default.
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