Identification of domains in an Arabidopsis acyl carrier protein gene promoter required for maximal organ-specific expression.
Deletions were made in the promoter of the acyl carrier protein (ACP) Acll.2 gene from Arabidopsis to investigate the nature of the cis-acting elements that direct its expression. These constructs, which included the untranslated leader region, were fused to a reporter gene coding for beta-glucuronidase (GUS) and transformed into tobacco. Quantitative fluorometric analysis of GUS activity in transgenic plants showed that expression in young leaves drops to a basal level when a 85 bp domain, from -320 to -236 relative to transcription initiation, is deleted. Maximum promoter activity in roots also depends on this domain, but two other regions are also important. In total, deletion of the sequences from -466 to -55 caused an ca. 80-fold reduction in Acl1.2 promoter activity in roots. The -320 to -236 domain forms a complex with a protein factor found in leaves and roots, which was not detectable in seeds. The formation of this protein-DNA complex was abolished by mutation of a bZIP core motif, ACGT, found within the context AAGACGTAG, which is dissimilar to the other bZIP-binding sites thus far characterized in plants. Previously we showed that Acl1.2 promoter activity is highest in seeds [2]. Here we find, in contrast to leaves and roots, that deletion to position -236 has no effect on GUS levels in seeds. However, nearly a 100-fold drop was observed when the -235 to -55 region was removed. Hence, this 180 bp domain contains all the cis-acting information necessary for Acl1.2 promoter activity in seeds. The same region is necessary for Acl1.2 activity in the receptacle, stigma, tapetum and pollen of the flower, as demonstrated by histochemical staining.
Baerson
S R
SR
Department of Molecular Genetics and Cell Biology, University of Chicago, IL 60637.
Vander Heiden
M G
MG
Lamppa
G K
GK
D000213:Acyl Carrier Protein / Q000235:genetics*; D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000302:isolation & purification; D051790:G-Box Binding Factors; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D017930:Genes, Reporter; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D012098:Reproduction; D017384:Sequence Deletion; D014018:Tissue Distribution; D014026:Nicotiana / Q000235:genetics; D014157:Transcription Factors
D000213:Acyl Carrier Protein; D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D051790:G-Box Binding Factors; D010940:Plant Proteins; D014157:Transcription Factors; C071572:acyl carrier proteins, barley