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Molecular and General Genetics MGG

, Volume 232, Issue 3, pp 383–393 | Cite as

Regulation and interaction of multiple protein factors with the proximal promoter regions of a rice high pl α-amylase gene

  • Ju-Kon Kim
  • Jun Cao
  • Ray Wu
Article

Summary

The α-amylase gene is known to be regulated by the plant hormone gibberellin (GA) in cereal aleurone cells. The accumulation of the mRNA corresponding to a rice high pl α-amylase gene, OSamy-c, was stimulated 20-fold by exogenous GA, in half-seeds lacking embryos. Regulatory regions in the promoter of this high pl subfamily were analyzed. The OSamy-c 5′ flanking sequence, spanning positions — 231 to + 29, was fused upstream of the β-glucuronidase (GUS) gene coding region. The delivery of this plasmid into rice aleurone cells by the biolistic method resulted in a GA-stimulated synthesis of GUS. Gel retardation assays were performed to study protein-DNA interactions between putative regulatory sequences of OSamy-c and partially purified rice seed extracts. We identified multiple seed-specific protein factors that bind to proximal regions of the OSamy-c promoter between positions — 231 and — 162. Five different proteins were distinguished based on competitive binding studies. Three protein binding regions were located by footprinting analyses, one of which is located in the conserved sequence also found upstream of other GA-inducible genes. Two protein factors in rice aleurone cells that interact with the putative regulatory sequence do not require GA induction.

Key words

Rice aleurone α-Amylase promoter Gibberellin regulation DNA-protein interaction Footprinting 

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Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • Ju-Kon Kim
    • 1
  • Jun Cao
    • 2
  • Ray Wu
    • 1
    • 3
  1. 1.Field of BotanyCornell UniversityIthacaUSA
  2. 2.Plant Science CenterCornell UniversityIthacaUSA
  3. 3.Section of Biochemistry, Molecular and Cell Biology, Division of Biological SciencesCornell UniversityIthacaUSA

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