Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1997 Feb 1;321(Pt 3):645–653. doi: 10.1042/bj3210645

Transcriptional regulatory elements in the upstream and intron of the fibroin gene bind three specific factors POU-M1, Bm Fkh and FMBP-1.

S Takiya 1, H Kokubo 1, Y Suzuki 1
PMCID: PMC1218118  PMID: 9032449

Abstract

The transcriptional modulator in the fibroin gene intron is composed of multiple octamer-like AT-rich elements, to which several specific DNA-binding proteins named fibroin-modulator-binding proteins (FMBPs) bind. Three major FMBPs in the silk gland were characterized. Two of them (FMBP-2 and -3) were identified as a Fork head homologue (Bm Fkh) and a POU-domain protein (POU-M1) respectively. These factors were expressed in the silk gland with distinct temporal- and spatial-specificities during late larval development as well as during embryogenesis, and did not correlate directly with fibroin gene expression. The other (FMBP-1) appeared to correlate with the expression of the fibroin gene for temporal- and spatial-specificity. These FMBPs also bind to the elements in the upstream modulator. Transcriptional enhancement by both modulators was inhibited by binding competition for these factors with oligonucleotides. These results suggest that expression of the fibroin gene is controlled by co-ordination of these factors with distinct specificities during silk-gland development.

Full Text

The Full Text of this article is available as a PDF (505.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Couble P., Chevillard M., Moine A., Ravel-Chapuis P., Prudhomme J. C. Structural organization of the P25 gene of Bombyx mori and comparative analysis of its 5' flanking DNA with that of the fibroin gene. Nucleic Acids Res. 1985 Mar 11;13(5):1801–1814. doi: 10.1093/nar/13.5.1801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Couble P., Michaille J. J., Garel A., Couble M. L., Prudhomme J. C. Developmental switches of sericin mRNA splicing in individual cells of Bombyx mori silkgland. Dev Biol. 1987 Dec;124(2):431–440. doi: 10.1016/0012-1606(87)90496-9. [DOI] [PubMed] [Google Scholar]
  3. Couble P., Moine A., Garel A., Prudhomme J. C. Developmental variations of a nonfibroin mRNA of Bombyx mori silkgland, encoding for a low-molecular-weight silk protein. Dev Biol. 1983 Jun;97(2):398–407. doi: 10.1016/0012-1606(83)90096-9. [DOI] [PubMed] [Google Scholar]
  4. Dirksen M. L., Jamrich M. A novel, activin-inducible, blastopore lip-specific gene of Xenopus laevis contains a fork head DNA-binding domain. Genes Dev. 1992 Apr;6(4):599–608. doi: 10.1101/gad.6.4.599. [DOI] [PubMed] [Google Scholar]
  5. Durand B., Drevet J., Couble P. P25 gene regulation in Bombyx mori silk gland: two promoter-binding factors have distinct tissue and developmental specificities. Mol Cell Biol. 1992 Dec;12(12):5768–5777. doi: 10.1128/mcb.12.12.5768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fukuta M., Matsuno K., Hui C. C., Nagata T., Takiya S., Xu P. X., Ueno K., Suzuki Y. Molecular cloning of a POU domain-containing factor involved in the regulation of the Bombyx sericin-1 gene. J Biol Chem. 1993 Sep 15;268(26):19471–19475. [PubMed] [Google Scholar]
  7. Hirose S., Suzuki Y. In vitro transcription of eukaryotic genes is affected differently by the degree of DNA supercoiling. Proc Natl Acad Sci U S A. 1988 Feb;85(3):718–722. doi: 10.1073/pnas.85.3.718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hui C. C., Matsuno K., Suzuki Y. Fibroin gene promoter contains a cluster of homeodomain binding sites that interact with three silk gland factors. J Mol Biol. 1990 Jun 20;213(4):651–670. doi: 10.1016/S0022-2836(05)80253-0. [DOI] [PubMed] [Google Scholar]
  9. Hui C. C., Suzuki Y. Enhancement of transcription from the Ad2 major late promoter by upstream elements of the fibroin- and sericin-1-encoding genes in silk gland extracts. Gene. 1989 Dec 28;85(2):403–411. doi: 10.1016/0378-1119(89)90433-2. [DOI] [PubMed] [Google Scholar]
  10. Jongens T. A., Fowler T., Shermoen A. W., Beckendorf S. K. Functional redundancy in the tissue-specific enhancer of the Drosophila Sgs-4 gene. EMBO J. 1988 Aug;7(8):2559–2567. doi: 10.1002/j.1460-2075.1988.tb03105.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lai E., Prezioso V. R., Smith E., Litvin O., Costa R. H., Darnell J. E., Jr HNF-3A, a hepatocyte-enriched transcription factor of novel structure is regulated transcriptionally. Genes Dev. 1990 Aug;4(8):1427–1436. doi: 10.1101/gad.4.8.1427. [DOI] [PubMed] [Google Scholar]
  12. Lai E., Prezioso V. R., Tao W. F., Chen W. S., Darnell J. E., Jr Hepatocyte nuclear factor 3 alpha belongs to a gene family in mammals that is homologous to the Drosophila homeotic gene fork head. Genes Dev. 1991 Mar;5(3):416–427. doi: 10.1101/gad.5.3.416. [DOI] [PubMed] [Google Scholar]
  13. Mach V., Takiya S., Ohno K., Handa H., Imai T., Suzuki Y. Silk gland factor-1 involved in the regulation of Bombyx sericin-1 gene contains fork head motif. J Biol Chem. 1995 Apr 21;270(16):9340–9346. doi: 10.1074/jbc.270.16.9340. [DOI] [PubMed] [Google Scholar]
  14. Maekawa H., Suzuki Y. Repeated turn-off and turn-on of fibroin gene transcription during silk gland development of Bombyx mori. Dev Biol. 1980 Aug;78(2):394–406. doi: 10.1016/0012-1606(80)90343-7. [DOI] [PubMed] [Google Scholar]
  15. Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Matsuno K., Hui C. C., Takiya S., Suzuki T., Ueno K., Suzuki Y. Transcription signals and protein binding sites for sericin gene transcription in vitro. J Biol Chem. 1989 Nov 5;264(31):18707–18713. [PubMed] [Google Scholar]
  17. Matsuno K., Takiya S., Hui C. C., Suzuki T., Fukuta M., Ueno K., Suzuki Y. Transcriptional stimulation via SC site of Bombyx sericin-1 gene through an interaction with a DNA binding protein SGF-3. Nucleic Acids Res. 1990 Apr 11;18(7):1853–1858. doi: 10.1093/nar/18.7.1853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Suzuki T., Matsuno K., Takiya S., Ohno K., Ueno K., Suzuki Y. Purification and characterization of an enhancer-binding protein of the fibroin gene. I. Complete purification of fibroin factor 1. J Biol Chem. 1991 Sep 5;266(25):16935–16941. [PubMed] [Google Scholar]
  19. Suzuki T., Takiya S., Matsuno K., Ohno K., Ueno K., Suzuki Y. Purification and characterization of an enhancer-binding protein of the fibroin gene. II. Functional analyses of fibroin factor 1. J Biol Chem. 1991 Sep 5;266(25):16942–16947. [PubMed] [Google Scholar]
  20. Suzuki Y. Genes that are involved in Bombyx body plan and silk gene regulation. Int J Dev Biol. 1994 Jun;38(2):231–235. [PubMed] [Google Scholar]
  21. Suzuki Y., Tsuda M., Hirose S., Takiya S. Transcription signals and factors of the silk genes. Adv Biophys. 1986;21:205–215. doi: 10.1016/0065-227x(86)90024-9. [DOI] [PubMed] [Google Scholar]
  22. Suzuki Y., Tsuda M., Takiya S., Hirose S., Suzuki E., Kameda M., Ninaki O. Tissue-specific transcription enhancement of the fibroin gene characterized by cell-free systems. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9522–9526. doi: 10.1073/pnas.83.24.9522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Takiya S., Hui C. C., Suzuki Y. A contribution of the core-promoter and its surrounding regions to the preferential transcription of the fibroin gene in posterior silk gland extracts. EMBO J. 1990 Feb;9(2):489–496. doi: 10.1002/j.1460-2075.1990.tb08135.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Takiya S., Suzuki Y. Factors involved in preferential transcription of the fibroin gene. Eur J Biochem. 1989 Jan 15;179(1):1–9. doi: 10.1111/j.1432-1033.1989.tb14513.x. [DOI] [PubMed] [Google Scholar]
  25. Tsuda M., Suzuki Y. Faithful transcription initiation of fibroin gene in a homologous cell-free system reveals an enhancing effect of 5' flanking sequence far upstream. Cell. 1981 Nov;27(1 Pt 2):175–182. doi: 10.1016/0092-8674(81)90371-8. [DOI] [PubMed] [Google Scholar]
  26. Tsuda M., Suzuki Y. Transcription modulation in vitro of the fibroin gene exerted by a 200-base-pair region upstream from the "TATA" box. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7442–7446. doi: 10.1073/pnas.80.24.7442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Weigel D., Jürgens G., Küttner F., Seifert E., Jäckle H. The homeotic gene fork head encodes a nuclear protein and is expressed in the terminal regions of the Drosophila embryo. Cell. 1989 May 19;57(4):645–658. doi: 10.1016/0092-8674(89)90133-5. [DOI] [PubMed] [Google Scholar]
  28. Xu P. X., Fukuta M., Takiya S., Matsuno K., Xu X., Suzuki Y. Promoter of the POU-M1/SGF-3 gene involved in the expression of Bombyx silk genes. J Biol Chem. 1994 Jan 28;269(4):2733–2742. [PubMed] [Google Scholar]
  29. Yamaguchi K., Kikuchi Y., Takagi T., Kikuchi A., Oyama F., Shimura K., Mizuno S. Primary structure of the silk fibroin light chain determined by cDNA sequencing and peptide analysis. J Mol Biol. 1989 Nov 5;210(1):127–139. doi: 10.1016/0022-2836(89)90295-7. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES