Sequence-specific interactions of a nuclear protein factor with the promoter region of a rice gene for α-amylase, RAmy3D

Shin Ichiro Mitsunaga, Raymond L. Rodriguez, Junji Yamaguchi

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

The expression of a rice gene for α-amylase, RAmy3D, in suspension-cultured cells is induced at the transcriptional level by the deprivation of sugars. Binding of a nuclear protein from suspension-cultured rice cells to the promoter region of the RAmy3D gene was studied by gel-retardation and DNase I footprinting assays. Gel-retardation assays indicated that a 358-bp fragment of the promoter region interacted specifically with a protein factor from suspension-cultured cells. DNase I footprinting analysis allowed us to define three protein-binding regions. Each of these protein-binding sequences contained the GCCG G/C CG motif, which is specifically present in the promoter region of the sugar-regulated gene, RAmy3D, for rice α-amylase and not in that of the gibberellin-regulated RAmy1A gene. Subsequent cross-competition experiments using gel-retardation assay and synthetic oligonucleotides showed that the GCCG G/C CG motifs directly mediated the binding of a nuclear protein. These observations are discussed in relation to expression of the gene for α-amylase in suspension-cultured cells.

Original languageEnglish (US)
Pages (from-to)1948-1953
Number of pages6
JournalNucleic Acids Research
Volume22
Issue number11
StatePublished - Jun 11 1994

Fingerprint

Amylases
Nuclear Proteins
Genetic Promoter Regions
Promoter
Cultured Cells
Suspensions
Genes
Gene
Protein
Deoxyribonuclease I
Electrophoretic Mobility Shift Assay
Assays
Interaction
Protein Binding
Gels
Cells
Sugars
Gibberellins
Gibberellin
Oligonucleotides

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Sequence-specific interactions of a nuclear protein factor with the promoter region of a rice gene for α-amylase, RAmy3D. / Mitsunaga, Shin Ichiro; Rodriguez, Raymond L.; Yamaguchi, Junji.

In: Nucleic Acids Research, Vol. 22, No. 11, 11.06.1994, p. 1948-1953.

Research output: Contribution to journalArticle

Mitsunaga, Shin Ichiro ; Rodriguez, Raymond L. ; Yamaguchi, Junji. / Sequence-specific interactions of a nuclear protein factor with the promoter region of a rice gene for α-amylase, RAmy3D. In: Nucleic Acids Research. 1994 ; Vol. 22, No. 11. pp. 1948-1953.
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