Multiprotein bridging factor 2 regulates the expression of the fibroin heavy chain gene by interacting with Bmdimmed in the silkworm Bombyx mori.

Abstract

Multiprotein bridging factor 2 (MBF2) was first isolated from the posterior silk gland of Bombyx mori. However, its function in B.mori is still unknown. Herein, MBF2 transcripts were detected mainly in the posterior silk gland and Malpighian tubules of B.mori larvae via a quantitative PCR analysis. An analysis of temporal expression patterns showed that the expression pattern of MBF2 was the opposite of that of the fibroin heavy chain (fibH) gene, as its expression was high during the fourth-instar moulting stage, decreased gradually during the fifth-instar feeding stage and disappeared at the end of the fifth-instar phase. Furthermore, bimolecular fluorescent complementation and Far-Western blot assays showed that MBF2 interacted with the basic helix-loop-helix transcription factor Bmdimmed. Dual luciferase reporter assays showed that MBF2 down-regulated the promoter activity of fibH and inhibited the effect of Bmdimmed (Bmdimm) on fibH expression. MBF2 expression was induced in silk glands after treatment with 20-hydroxyecdysone in vivo and in vitro. These findings suggest that MBF2 is a transcriptional repressor that is involved in controlling the regulation of the fibH gene in the posterior silk gland by interacting with Bmdimm.