Both Internal and External Regulatory Elements Control Expression of the Pea Fed-1 Gene in Transgenic Tobacco Seedlings

Gallo‐Meagher, Maria; Sowinski, Dolores A.; Elliott, Robert C.; Thompson, William F.

doi:10.2307/3869441

Cited by 13 publications

(16 citation statements)

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“…Photoregulatory elements have been found downstream of the transcription start site in other plant genes (23,24); however, it is not known whether these affect transcription or post-transcriptional events. Here we demonstrate that the presence of the rbcS-m3 3' end in the chimeric gene results in the photoregulated suppression of its transcription preferentially in MC.…”

Section: Lalizing the Region Of Rbcs-m3 Required For Enhancingmentioning

confidence: 99%

Transcriptional photoregulation of cell-type-preferred expression of maize rbcS-m3: 3' and 5' sequences are involved.

Viret

Mabrouk²,

Bogorad³

1994

Proc. Natl. Acad. Sci. U.S.A.

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In the C4 plant maize, members of the rbcS gene family, encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase, are not expressed in mesophyll cells (MC) but are expressed strongly in the adjacent bundle sheath cells (BSC). Expression of genes in an in situ transient expression assay indicates that the photostimulated expression seen in BSC during the first 24 h that leaves of dark-grown seedlings are illuminated requires rbcS-m3 sequences lying between -211 bp and +434 bp of the transcription start site. Photoregulated partial suppression of rbcS-m3 expression in MC, on the other hand, requires gene sequences that lie between -907 bp and -445 bp together with sequences that lie between +720 and +957 bp within the 3' transcribed region ofthe gene. Suppression in MC occurs during the second 24-h period that dark-grown seedlings have been illuminated, but not during the first 24 h. The 3' +720-to +957-bp region is also effective in lowering MC expression when it is relocated to a position >2 kbp upstream of the transcription start site. Thus, suppression of rbcS-m3 expression in MC has, at the least, a substantial transcriptional component. As reported earlier, a converse pattern of suppression in BSC and stimulation of expression in MC is seen in the control of cab-ml in maize leaves.In C4 plants, such (3,4), but within the first 24 h of illumination their abundance increases 2-to 3-fold in BSC and they are undetectable in MC (3, 4). Transcripts of the maize gene rbcS-m3 (5) follow this pattern of BSC-preferred accumulation upon illumination of dark-grown leaves and constitute about 35% of the total leaf rbcS mRNA in 24-h illuminated dark-grown maize (3). Using a 3-glucuronidase (GUS) in situ transient expression assay (6), we found that expression from a reporter gene containing 2.1 kbp upstream of the rbcS-m3 transcription start site [-2.1-kbp rbcS-m3 promoter (Pr):GUS:nopaline synthase (nos) terminator] is about the same in MC and BSC of 24-h illuminated leaves. GUS expression is promoted 2-to 3-fold in BSC by illumination, as expected from the behavior of rbcS-m3, but this reporter gene does not behave like rbcS-m3 in MC after illumination: GUS expression from the rbcS-m3 promoter is about the same in MC of unilluminated and illuminated leaves (3, 6).We have now found that inclusion of a 238-bp sequence from within the transcribed 3' end of rbcS-m3 (extending from +720 to +957 bp relative to the transcription start site) in a reporter gene containing 2.1 kbp of rbcS-m3 from upstream of the transcription start site preferentially reduces expression of GUS in MC in leaves of dark-grown seedlings that are greened (i.e., illuminated) for 24 h prior to bombardment. Furthermore, the 238-bp fragment provides MCpreferred suppression of expression through a transcriptional mechanism. Through a series of promoter deletion experi- (Fig. 1). The pMT211 plasmid was obtained by first digesting pMT444 with Kpn I and HindIII, then treating it with exonuclease III and mung bean nuclease, ...

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Section: Lalizing the Region Of Rbcs-m3 Required For Enhancingmentioning

confidence: 99%

Transcriptional photoregulation of cell-type-preferred expression of maize rbcS-m3: 3' and 5' sequences are involved.

Viret

Mabrouk²,

Bogorad³

1994

Proc. Natl. Acad. Sci. U.S.A.

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“…The membranes were then washed in a 0.1ϫ SSC and 0.1% (w/v) SDS buffer at 68°C. For control of RNA loading, membranes were rehybridized with an oligonucleotide probe complementary to 18S rRNA (Gallo-Meagher et al, 1992). The signals were visualized using a bio-imaging analyzer (BAS 2000, Fuji, Japan).…”

Section: Rna Extraction and Gel-blot Analysismentioning

confidence: 99%

The First Step of Gibberellin Biosynthesis in Pumpkin Is Catalyzed by at Least Two Copalyl Diphosphate Synthases Encoded by Differentially Regulated Genes

Smith

Yamaguchi²,

Aït-Ali³

et al. 1998

Plant Physiology

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The first step in gibberellin biosynthesis is catalyzed by copalyl diphosphate synthase (CPS) and ent-kaurene synthase. We have cloned from pumpkin (Cucurbita maxima L.) two cDNAs, CmCPS1 and CmCPS2, that each encode a CPS. Both recombinant fusion CmCPS proteins were active in vitro. CPS are translocated into plastids and processed by cleavage of transit peptides. For CmCPS1 and CmCPS2, the putative transit peptides cannot exceed the first 99 and 107 amino acids, respectively, because longer N-terminal deletions abolished activity. Levels of both CmCPS transcripts were strictly regulated in an organ-specific and developmental manner. Both transcripts were almost undetectable in leaves and were abundant in petioles. CmCPS1 transcript levels were high in young cotyledons and low in roots. In contrast, CmCPS2 transcripts were undetectable in cotyledons but present at significant levels in roots. In hypocotyls, apices, and petioles, CmCPS1 transcript levels decreased with age much more rapidly than those of CmCPS2. We speculate that CmCPS1 expression is correlated with the early stages of organ development, whereas CmCPS2 expression is correlated with subsequent growth. In contrast, C. maxima ent-kaurene synthase transcripts were detected in every organ at almost constant levels. Thus, ent-kaurene biosynthesis may be regulated through control of CPS expression.

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“…Ferredoxin-1 (Fed-1), a nuclear gene from pea encoding the major chloroplast isoform of ferredoxin, exemplifies this complexity. Fed-1 mRNA abundance is regulated by light at the level of transcription initiation in etiolated seedlings (Gallo-Meagher et al, 1992). In green leaves, however, Fed-1 mRNA abundance is regulated primarily by changes in mRNA stability (Elliot et al, 1989;Petracek et al, 1998b).…”

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confidence: 99%

The 5′ End of the Pea Ferredoxin-1 mRNA Mediates Rapid and Reversible Light-Directed Changes in Translation in Tobacco

Hansen

Petracek

Dickey³

et al. 2001

Plant Physiology

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Ferredoxin-1 (Fed-1) mRNA contains an internal light response element (iLRE) that destabilizes mRNA when light-grown plants are placed in darkness. mRNAs containing this element dissociate from polyribosomes in the leaves of transgenic tobacco (Nicotiana tabacum) plants transferred to the dark for 2 d. Here, we report in vivo labeling experiments with a chloramphenicol acetyl transferase mRNA fused to the Fed-1 iLRE. Our data indicate that the Fed-1 iLRE mediates a rapid decline in translational efficiency and that iLRE-containing mRNAs dissociate from polyribosomes within 20 min after plants are transferred to darkness. Both events occur before the decline in mRNA abundance, and polyribosome association is rapidly reversible if plants are re-illuminated. These observations support a model in which Fed-1 mRNA in illuminated leaves is stabilized by its association with polyribosomes, and/or by translation. In darkness a large portion of the mRNA dissociates from polyribosomes and is subsequently degraded. We also show that a significant portion of total tobacco leaf mRNA is shifted from polyribosomal to non-polyribosomal fractions after 20 min in the dark, indicating that translation of other mRNAs is also rapidly down-regulated in response to darkness. This class includes some, but not all, cytoplasmic mRNAs encoding proteins involved in photosynthesis.

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Both Internal and External Regulatory Elements Control Expression of the Pea Fed-1 Gene in Transgenic Tobacco Seedlings

Cited by 13 publications

References 0 publications

Transcriptional photoregulation of cell-type-preferred expression of maize rbcS-m3: 3' and 5' sequences are involved.

Transcriptional photoregulation of cell-type-preferred expression of maize rbcS-m3: 3' and 5' sequences are involved.

The First Step of Gibberellin Biosynthesis in Pumpkin Is Catalyzed by at Least Two Copalyl Diphosphate Synthases Encoded by Differentially Regulated Genes

The 5′ End of the Pea Ferredoxin-1 mRNA Mediates Rapid and Reversible Light-Directed Changes in Translation in Tobacco

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