The SebHLH transcription factor mediates trans-activation of the SeFAD2 gene promoter through binding to E- and G-box elements

Mi Jung Kim, Jeong Kook Kim, Jeong Sheop Shin, Mi Chung Suh

Research output: Contribution to journalArticlepeer-review

37 Citations (Scopus)

Abstract

Microsomal oleic acid desaturase (FAD2) catalyzes the first extra-plastidial desaturation in plants, converting oleic acid to linoleic acid, which is a major constituent in all cellular membranes as well as in seed storage oils. Seed-specific FAD2 (SeFAD2) produced 40% of linoleic acids in the total fatty acids of sesame (Sesamum indicum) seeds. The expression of SeFAD2 transcripts was spatially and temporally controlled during seed development. To investigate the regulatory mechanism controlling seed-specific SeFAD2 expression, we isolated a well-matched sequence homologous to the basic region/helix-loop-helix proteins by yeast one-hybrid screening and named it SebHLH. SebHLH transcripts were expressed in developing seeds and roots of sesame. SebHLH:GFP fusion protein localized in the nucleus. Recombinant SebHLH protein bound E-box (CANNTG) and G-box (CACGTG) elements in the region from -179 to -53 of the SeFAD2 gene promoter, and the external C and G nucleotides in the E- and G-box motifs were essential for SebHLH protein binding. The SebHLH gene, under the CaMV35S promoter, and the GUS reporter gene driven by E- and G-box motifs were co-expressed in developing sesame seeds and Arabidopsis transgenic leaves. This co-expression demonstrated that SebHLH protein mediates transactivation of the SeFAD2 gene promoter through binding to E- and G-box elements. E- or G-box elements frequently occur in the 5′-flanking region of genes that are involved in triacylglycerol biosynthesis and that exhibit seed-specific expression in Arabidopsis and other plants, suggesting that bHLH transcription factors play a key role in the transcriptional regulation of genes related to storage lipid biosynthesis and accumulation during seed development.

Original languageEnglish
Pages (from-to)453-466
Number of pages14
JournalPlant Molecular Biology
Volume64
Issue number4
DOIs
Publication statusPublished - 2007 Jul

Bibliographical note

Funding Information:
Acknowledgements The authors would like to thank Kyung Hee Paek for providing the pCAMBIA 1305.1 vector containing the minimal CaMV35S promoter and the NOS terminator and Sung Han Ok for helpful discussion about yeast one-hybrid screening. This work was supported by grants from the Agricultural Plant Stress Research Center (R11-2001-0920301-0) and the Interdisciplinary Research Program (R01-2006-000-11056-0) of the Korea Science and Engineering Foundation.

Keywords

  • FAD2
  • Microsomal oleic acid desaturase
  • SebHLH
  • Sesame
  • Sesamum indicum
  • bHLH transcription factor

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Genetics
  • Plant Science

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