Basic helix-loop-helix transcription factor Bmsage is involved in regulation of fibroin H-chain gene via interaction with SGF1 in Bombyx mori

PLoS One. 2014 Apr 16;9(4):e94091. doi: 10.1371/journal.pone.0094091. eCollection 2014.

Abstract

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 Bmsage; 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 4th instar molting stages, which increases gradually in the 5th 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 5th 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Basic Helix-Loop-Helix Transcription Factors / physiology*
  • Bombyx / genetics*
  • Fibroins / genetics*
  • Fibroins / metabolism
  • Gene Expression Regulation*
  • Insect Proteins / genetics
  • Insect Proteins / metabolism
  • Insect Proteins / physiology*
  • Molecular Sequence Data
  • Sequence Alignment
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Trans-Activators / physiology

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • Insect Proteins
  • Trans-Activators
  • silk gland factor-1
  • Fibroins

Grants and funding

This work was supported by grants from the National Hi-Tech Research and Development Program of China (No. 2011AA100306 to Q-YX), Program for New Century Excellent Talents (No. NCET-11-0699 to CL), the National Natural Science Foundation of China (No. 31000545 and No. 31372380 to CL) and Chongqing City Board of Education¼∧No. CY120101 to CL). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.