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The 5′ UTR negatively regulates quantitative and spatial expression from the ABI3 promoter

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Abstract

The involvement of transcription factors Arabidopsis abscisic acid-insensitive3 (ABI3), maize viviparous1 (VP1) and Phaseolus vulgaris ABI3-like factor (PvALF) in the spatial control of storage protein gene expression is well established. However, little insight exists as to how they are themselves regulated. To address this, a 5.15 kb ABI3 upstream sequence including a 4.6 kb full-length promoter and 519 bp of 5′-untranslated region (UTR) was used to drive either β-glucuronidase (GUS) or green fluorescent protein (GFP) expression in Arabidopsis. Expression from the full-length (−4630/+519ABI3) and various 5′-truncated promoters was detected during embryogenesis in all lines, except those transgenic for promoter elements shorter than 364 bp. Two upstream activating regions, −3600 to −2033 and −2033 to −882, enhanced GUS expression in seeds. The −882 to −364 region was sufficient to confer seed-specific expression of GUS when fused to a −64/+6CaMV 35S minimal promoter. Expression from the ABI3 promoter constructs was seed-specific, except in the presence of exogenous abscisic acid (ABA) (>0.3 μM), when GUS expression was detected in seedling roots. Excision of a 405 bp region containing three upstream open reading frames (uORFs) from the 5′-UTR dramatically increased GUS expression and debilitated constraint of reporter expression in roots. Negative regulation of ABI3 expression by the 5′-UTR may involve a post-transcriptional mechanism analogous to that of tumor suppressor genes which also bear long, uORF-containing, 5′-UTRs, or through interactions with RNA-binding proteins.

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  1. Institute of Developmental and Molecular Biology and Department of Biology, Texas A&M University, College Station, TX, 77843-3155, USA

    Danny W-K Ng, Mahesh B. Chandrasekharan & Timothy C. Hall

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  1. Danny W-K Ng
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Correspondence to Timothy C. Hall.

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Ng, D.WK., Chandrasekharan, M.B. & Hall, T.C. The 5′ UTR negatively regulates quantitative and spatial expression from the ABI3 promoter. Plant Mol Biol 54, 25–38 (2004). https://doi.org/10.1023/B:PLAN.0000028767.06820.34

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