Analysis of cis-regulatory elements involved in the activation of a member of chalcone synthase gene family (PsCHs1) in pea

Hikaru Seki, Yuki Ichinose, Hisaharu Kato, Tomonori Shiraishi, Tetsuji Yamada

Research output: Contribution to journalArticle

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Abstract

Cis-regulatory elements involved in the activation of the plant defense- related gene encoding chalcone synthase 1 (PsChs1) in pea (Pisum sativum L.) were examined by transient transfection, gel mobility shift assay and in vitro DNase I-footprinting analysis. Transient transfection assay revealed that a 61 bp DNA fragment spanning from -242 to -182 of PsChs1 was required for the maximal promoter activity and possibly involved in the enhancement of elicitor-mediated activation. Nuclear isolate from elicitor-treated pea epicotyl tissues contained some factor(s) that specifically bound to this DNA fragment to form a complex with low mobility (LMC, low mobility complex) in gel mobility shift assay. DNase I-footprinting analysis of LMC revealed that among three protected regions detected in a 61 bp DNA fragment, two regions contained identical AT-rich sequence, TAAAATACT. Site directed mutation in either or both identical sequences, TAAAATACT to TGGAATACT, resulted in the reduction or loss in the ability to form LMC. Detailed analysis of 61 bp DNA fragment demonstrated that the region from -242 to -226 containing promoter- distal TAAAATACT motif was imperative for the maximal elicitor-mediated activation of PsChs1.

Original languageEnglish
Pages (from-to)479-491
Number of pages13
JournalPlant Molecular Biology
Volume31
Issue number3
Publication statusPublished - Jun 1996

Fingerprint

regulatory sequences
naringenin-chalcone synthase
Peas
peas
Genes
Chemical activation
deoxyribonuclease I
Assays
Deoxyribonuclease I
DNA
Electrophoretic Mobility Shift Assay
transfection
Transfection
assays
genes
Gels
promoter regions
AT Rich Sequence
gels
Gene encoding

Keywords

  • AT-rich sequences
  • chalcone synthase
  • elicitor
  • low mobility complex (LMC)
  • Mycosphaerella pinodes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Analysis of cis-regulatory elements involved in the activation of a member of chalcone synthase gene family (PsCHs1) in pea. / Seki, Hikaru; Ichinose, Yuki; Kato, Hisaharu; Shiraishi, Tomonori; Yamada, Tetsuji.

In: Plant Molecular Biology, Vol. 31, No. 3, 06.1996, p. 479-491.

Research output: Contribution to journalArticle

Seki, Hikaru ; Ichinose, Yuki ; Kato, Hisaharu ; Shiraishi, Tomonori ; Yamada, Tetsuji. / Analysis of cis-regulatory elements involved in the activation of a member of chalcone synthase gene family (PsCHs1) in pea. In: Plant Molecular Biology. 1996 ; Vol. 31, No. 3. pp. 479-491.
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