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F Rook

Publications and source records attributed to F Rook.

7 recordsLinked to original sources
PubMed2001

Impaired sucrose-induction mutants reveal the modulation of sugar-induced starch biosynthetic gene expression by abscisic acid signalling.

Plants both produce and utilize carbohydrates and have developed mechanisms to regulate their sugar status and co-ordinate carbohydrate partitioning. High sugar levels result in a feedback inhibition of photosynthesis and an induction of storage processes. We used a genetic approach to isolate components of the signalling pathway regulating the induction of starch biosynthesis. The regulatory sequences of the sugar inducible ADP-glucose pyrophosphorylase subunit ApL3 were fused to a negative selection marker. Of the four impaired sucrose induction (isi) mutants described here, two (isi1 and isi2) were specific to this screen. The other two mutants (isi3 and isi4) showed additional phenotypes associated with sugar-sensing screens that select for seedling establishment on high-sugar media. The isi3 and isi4 mutants were found to be involved in the abscisic acid signalling pathway. isi3 is allelic to abscisic acid insensitive4 (abi4), a gene encoding an Apetala2-type transcription factor; isi4 was found to be allelic to glucose insensitive1 (gin1) previously reported to reveal cross-talk between ethylene and glucose signalling. Here we present an alternative interpretation of gin1 as an allele of the ABA-deficient mutant aba2. Expression analysis showed that ABA is unable to induce ApL3 gene expression by itself, but greatly enhances ApL3 induction by sugar. Our data suggest a major role for ABA in relation to sugar-signalling pathways, in that it enhances the ability of tissues to respond to subsequent sugar signals.

Abscisic Acid↗
PubMed1998

The light-regulated Arabidopsis bZIP transcription factor gene ATB2 encodes a protein with an unusually long leucine zipper domain.

A light-regulated basic domain/leucine zipper gene, ATB2, was identified in an Arabidopsis thaliana transcription factor gene collection. Both genomic and cDNA clones of ATB2 were isolated. The gene encodes a small protein (18 kDa) which mainly consists of the basic domain and an unusually long leucine zipper. The expression of the ATB2 gene is induced when etiolated or dark-adapted seedlings are transferred to the light. Moreover, its expression is derepressed in dark-grown seedlings of the photomorphogenic mutants cop1 and det1. In mature plants, transcript levels are particularly high in flowers and also light-responsive in these tissues.

Adaptation, Biological↗
PubMed1998

Sucrose-specific signalling represses translation of the Arabidopsis ATB2 bZIP transcription factor gene.

The Arabidopsis bZIP transcription factor gene ATB2 has been shown previously to be expressed in a light-regulated and tissue-specific way. Here we describe the precise localization of ATB2 expression, using transgenic lines containing an ATB2 promoter-GUS reporter gene construct. The observed expression pattern suggests a role for ATB2 in the control of processes associated with the transport or utilization of metabolites. Remarkably, expression of the ATB2-GUS reporter gene construct was specifically repressed by sucrose. Other sugars, such as glucose and fructose, alone or in combination, were ineffective. Repression was observed at external sucrose concentrations exceeding 25 mM. Transcript levels of both the endogenous ATB2 gene and the ATB2-GUS reporter gene were not repressed by sucrose, suggesting that sucrose affects mRNA translation. This translational regulation involves the ATB2 leader sequence because deletion of the leader resulted in loss of sucrose repression. Our results provide evidence for a sucrose-specific sugar sensing and signalling system in plants.

Arabidopsis↗
PubMed1995

The homeobox gene ATH1 of Arabidopsis is derepressed in the photomorphogenic mutants cop1 and det1.

A light-regulated Arabidopsis thaliana homeobox 1 gene (ATH1) was identified in a transcription factor gene collection. Primer extension analysis of this gene showed the presence of two major transcripts with unusually long 5' untranslated leader sequences. The leader sequence of the isolated cDNA clone contains several small open reading frames upstream of the initiation codon of the largest open reading frame coding for the homeodomain protein. This leader sequence was found to affect the translation efficiency negatively in an in vitro translation system. The expression of the ATH1 gene is dependent on the presence of light. After illuminating etiolated or dark-adapted Arabidopsis seedlings, the ATH1 mRNA level increased rapidly. Expression of ATH1 does not require the presence of active chloroplasts because photooxidative destruction of the chloroplast by norflurazon treatment did not influence the ATH1 mRNA level. In dark-grown seedlings of the photomorphogenic constitutive photomorphogenic 1 (cop1) and deetiolated 1 (det1) mutants, the ATH1 mRNA level was elevated. This shows that the gene products of these loci directly or indirectly repress ATH1 expression in etiolated wild-type seedlings. A correlation between the strength of the cop1 allele and the ATH1 mRNA level was found. This relationship suggests a role for the ATH1 protein in the signal transduction pathway downstream of COP1.

Amino Acid Sequence↗
PubMed1994

Methotrexate does not block import of a DHFR fusion protein into chloroplasts.

Protein import into chloroplasts requires the movement of a precursor protein across the envelope membranes. The conformation of a precursor as it passes from the aqueous medium across the hydrophobic membranes is not known in detail. To address this problem we examined precursor conformation during translocation using the chimeric precursor PCDHFR, which contains the plastocyanin (PC) transit peptide in front of mouse cytosolic dihydrofolate reductase (DHFR). The chimeric protein is targeted to chloroplasts and is competent for import. The conformation of PCDHFR can be stabilized by complexing with methotrexate, an analogue of the substrate of DHFR. Methotrexate strongly inhibits DHFR import into yeast mitochondria (M. Eilers and G. Schatz, Nature 322 (1986) 228-232), presumably because the precursor must unfold to cross the membrane and it cannot do so when complexed with methotrexate. We show here that methotrexate does not block PCDHFR import into chloroplasts. Methotrexate does slow the rate of import, and protects DHFR from degradation once inside chloroplasts. The processed protein is localized in the stroma, indicating that import into thylakoids is impeded. Protease sensitivity assays indicate that the complex of precursor protein with methotrexate changes in conformation during the translocation across the envelope.

Amino Acid Sequence↗