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Silk glands are specialized in the synthesis of several secretory proteins. Expression of genes encoding the silk proteins in  Bombyx mori  silk glands with strict territorial and developmental specificities is regulated by many transcription factors. In this study, we have characterized  B. mori sage , which is closely related to  sage  in the fruitfly  Drosophila melanogaster . It is termed  Bmsag e; it encodes transcription factor Bmsage, which belongs to the Mesp subfamily, containing a basic helix–loop–helix motif. Bmsage transcripts were detected specifically in the silk glands of  B. mori  larvae through RT-PCR analysis. Immunoblotting analysis confirmed the Bmsage protein existed exclusively in  B. mori  middle and posterior silk gland cells.  Bmsage  has a low level of expression in the 4 th  instar molting stages, which increases gradually in the 5 th  instar feeding stages and then declines from the wandering to the pupation stages. Quantitative PCR analysis suggested the expression level of  Bmsage  in a high silk strain was higher compared to a lower silk strain on day 3 of the larval 5 th  instar. Furthermore, far western blotting and co-immunoprecipitation assays showed the Bmsage protein interacted with the fork head transcription factor silk gland factor 1 (SGF1). An electrophoretic mobility shift assay showed the complex of Bmsage and SGF1 proteins bound to the A and B elements in the promoter of  fibroin H-chain  gene (fib-H) , respectively. Luciferase reporter gene assays confirmed the complex of Bmsage and SGF1 proteins increased the expression of  fib-H . Together, these results suggest Bmsage is involved in the regulation of the expression of  fib-H  by being together with SGF1 in  B. mori  PSG cells.

Basic Helix-Loop-Helix Transcription Factor Bmsage Is Involved in Regulation of fibroin H-chain Gene via Interaction with SGF1 in Bombyx mori