Functional Analysis of Linker Insertions and Point Mutations in the Alpha-Amy2/54 GA-regulated Promoter

Overview

Journal Plant Mol Biol

Date 1995 Nov 1

PMID 8541501

Citations 3

Authors

J W Tregear

L F Primavesi

A K Huttly

Affiliations

Soon will be listed here.

Abstract

Functional analysis of a gibberellin-regulated wheat alpha-amylase promoter, alpha-Amy2/54, has indicated that three regions were essential for expression. By studying the ability of mutant promoters, containing a randomly inserted 22 bp excision linker, to direct expression in oat aleurone protoplasts we have refined the positions and extents of these three cis elements and also demonstrated the presence of two additional elements. By converting the linker insertions to either single base point mutations or deletions using the class IIS restriction endonuclease BsmI we have shown that nucleotides -119 and -109 within the GARE -121GTAACAGAGTCTGG-108 and nucleotide -152 within the proposed element -156GATTGACTTGACC-144 are essential for high level expression from this promoter.

Citing Articles

Heme oxygenase-1 is involved in nitric oxide- and cGMP-induced α-Amy2/54 gene expression in GA-treated wheat aleurone layers.

Wu M, Wang F, Zhang C, Xie Y, Han B, Huang J Plant Mol Biol. 2012; 81(1-2):27-40.

PMID: 23090695 DOI: 10.1007/s11103-012-9979-x.

DNase1 footprints suggest the involvement of at least three types of transcription factors in the regulation of alpha-Amy2/A by gibberellin.

Willmott R, Rushton P, Hooley R, Lazarus C Plant Mol Biol. 1998; 38(5):817-25.

PMID: 9862499 DOI: 10.1023/a:1006084104041.

Members of a new family of DNA-binding proteins bind to a conserved cis-element in the promoters of alpha-Amy2 genes.

Rushton P, Macdonald H, Huttly A, Lazarus C, Hooley R Plant Mol Biol. 1995; 29(4):691-702.

PMID: 8541496 DOI: 10.1007/BF00041160.

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