Regulation of HEMA1 expression by phytochrome and a plastid signal during de-etiolation in Arabidopsis thaliana.
The synthesis of 5-aminolevulinic acid (ALA) is the rate-limiting step for the formation of all plant tetrapyrroles, including chlorophyll and heme, and regulation of ALA synthesis is therefore critical to plant development. Glutamyl-tRNA reductase (GluTR) is the first committed enzyme of this pathway and is encoded by a small family of nuclear HEMA genes. Here, we have used transgenic Arabidopsis (Arabidopsis thaliana L. Col) lines expressing chimeric HEMA1 promoter:gusA fusion genes, combined with RNA gel blot analyses, to characterise the light-mediated regulation of the Arabidopsis HEMA1 gene during de-etiolation. HEMA1 was expressed strongly, but not exclusively, in photosynthetic tissues and was shown to be light regulated at the transcriptional level by the phytochrome family of photoreceptors acting in both the far-red high irradiance and low fluence response modes. The HEMA2 gene, which is expressed only in roots of seedlings, was not light regulated. Analysis of truncated HEMA1 promoter constructs demonstrated that a -199/+252 promoter fragment was sufficient to confer full light-responsiveness to gusA expression. This fragment contained GT-1/I-box and CCA-1 binding sites that are implicated as the light-responsive cis elements. Both the full-length and truncated HEMA1 promoters required the presence of intact chloroplasts for full expression, consistent with previous indications that light and plastid factor signals converge to co-ordinately regulate expression of photosynthesis-related nuclear genes. These results provide the most comprehensive analysis to date of the light-regulation of a tetrapyrrole biosynthetic gene and support a direct link between regulation of HEMA1 transcription and chlorophyll accumulation during seedling de-etiolation.
McCormac
A C
AC
School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK.
Fischer
A
A
Kumar
A M
AM
Söll
D
D
Terry
M J
MJ
D000445:Aldehyde Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D000622:Aminolevulinic Acid / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D015152:Blotting, Northern; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D020723:Glucuronic Acid / Q000378:metabolism; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D017440:Photoperiod; D010834:Phytochrome / Q000378:metabolism*; D018087:Plastids / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D018749:RNA, Plant / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D018749:RNA, Plant; D011993:Recombinant Fusion Proteins; D010834:Phytochrome; D000622:Aminolevulinic Acid; D020723:Glucuronic Acid; D000445:Aldehyde Oxidoreductases; C067282:glutamyl tRNA reductase