An enhancer-like element present in the promoter of a T-DNA gene from the Ti plasmid of
            <i>Agrobacterium tumefaciens</i>

Bruce, William A.; Bandyopadhyay, Ram; Gurley, William B.

doi:10.1073/pnas.85.12.4310

Cited by 15 publications

(14 citation statements)

References 33 publications

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“…Enhancer-like sequences from the T-DNA 780 gene [4,5], a pentamer of an AT-rich region from a soybean fi-conglycinin gene [7,10] and a trimer of an AT-composite sequence from a soybean heat shock gene [9] were cloned into the Barn HI site of the -325 5' deletion after addition ofBgl II linkers. PCR was then used to produce fragments containing the enhancer sequences and ocs motif that were cloned into the 5' deletion constructs.…”

Section: Construction Of Plasmids Containing Mas Gene Deletionsmentioning

confidence: 99%

“…We are interested in the structure of rnas and other T-DNA genes [4,5,44] and in improved expression of selectable markers for transformation of the Grarnineae [21,40]. In this paper we further delineate the areas of the rnas gene essential for transcription and attempt to enhance transcriptional efficacy of the rnas promoter in maize protoplasts.…”

Section: Introductionmentioning

confidence: 99%

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Multiple ocs-like elements required for efficient transcription of the mannopine synthase gene of T-DNA in maize protoplasts

Fox

Vasil

et al. 1992

Plant Mol Biol

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Regulatory elements controlling transcriptional activity of the mannopine synthase 2' promoter (mas 2') were defined by analysis of deletion mutants in transient expression assays in maize protoplasts. Deletion of the region between -305 and -290 containing sequence similarity to the octopine synthase (ocs) promoter element reduced activity by 67% compared to wild type activity. Less than 1% of the activity remained in 5' deletions downstream of -153. Inclusion of various heterologous enhancer-like sequences immediately upstream of position -325 increased activity by up to 7.5-fold. Insertion of the -325 to -275 sequence alone, or in combination with heterologous enhancer-like elements, restored activity of some of the 5'-deletion mutants. Restoration of activity was not obtained with mutants deleted past position -127. Our results suggest that a single class of nuclear proteins from maize interact with high affinity at elements designated mas b (-306 to -275; mas 1' element), d (-127 to -108), and e (-82 to -39; mas 2' element) as well as the 20 bp element from the ocs promoter. Although the binding site at mas d only appears to accommodate a single protein, this element has the potential to make a weak, but positive, contribution to the activity of the mas 2' promoter. The binding of nuclear proteins could not be demonstrated at mas a and c, both of which showed limited homology to the ocs element. Mutational evidence suggested that mas a and c may also contribute to mas 2' transcription.

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Section: Construction Of Plasmids Containing Mas Gene Deletionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Multiple ocs-like elements required for efficient transcription of the mannopine synthase gene of T-DNA in maize protoplasts

Fox

Vasil

et al. 1992

Plant Mol Biol

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“…This transcription factor also binds to the regulatory elements of cauliflower mosaic virus (CaMV) 35S and nopaline synthase (nos) promoters, suggesting the presence of a common regulatory sequence. Analyses of promoters from other T-DNA genes such as gene 5 [27], 780 [10], agropine biosynthase [10], mannopine synthase [ 14,31], and isopentenyl transferase [ 13,37] showed that an ocs-like element is also present in these promoters.…”

Section: Introductionmentioning

confidence: 99%

A 20 nucleotide upstream element is essential for the nopaline synthase (nos) promoter activity

et al. 1994

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The nopaline synthase (nos) promoter is expressed in a wide range of plant cell types and regulated by various developmental and environmental factors. The nos upstream control region essential for this regulation was studied by means of synthetic oligomers using transient and stable transformation systems. Insertion of a 20 nucleotide sequence containing two hexamer motifs and a spacer region into deletion mutants lacking the upstream control region was essential for promoter activity. Mutation of one or more nucleotides of either hexamer sequence significantly altered the strength of expression of the nos promoter. Point mutations within the spacer region also strongly influenced promoter strength. Insertion of multiple copies of the 20 nucleotide sequence into the nonfunctional deletion mutants proportionally increased the promoter activity. These results suggest that this twenty nucleotide sequence is essential for the nos promoter to function. Substitution of the nos element with the ocs or 35S as-1 which contain similar hexamer motifs restored not only promoter activity but also responses to wounding, auxin, methyl jasmonate, and salicylic acid.

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“…Initial studies by Bruce et al [9,10] using a 5' deletion approach revealed the existence of a region from -440 to -229 (designated as the activator or enhancer) that was responsible for 99~ of promoter activity when assayed in sunflower tumors. When tested for enhancer-like activity using a -37 5' deletion of the 780 gene as a reporter, the activator region was found to strongly stimulate gene expression in both orientations when positioned upstream of TATA at either a proximal or distal location.…”

Section: Introductionmentioning

confidence: 99%

Site-directed mutagenesis of the enhancer region of the 780 gene promoter of T-DNA

O’Grady

Gurley

1995

Plant Mol Biol

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Potential regulatory sequences within the enhancer-like region of the 780 gene promoter (Agrobacterium tumefaciens T-DNA) were identified by site-directed mutagenesis. Transcriptional activity of the mutated promoter was analyzed by S1 nuclease mapping of RNA from crown gall tumors of sunflower incited using a T-DNA-based vector. Variability in expression levels were minimized by the use of an internal reference gene and the pooling of at least 200 tumors per construct tested. This approach identified numerous sequences that influence transcriptional activity in either a positive or negative manner. Eight regions of positive influence and three of negative were identified from analysis of those mutations that exhibited low variability in expression (P < 0.005) and affected activity by at least 20%.

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An enhancer-like element present in the promoter of a T-DNA gene from the Ti plasmid of Agrobacterium tumefaciens

Cited by 15 publications

References 33 publications

Multiple ocs-like elements required for efficient transcription of the mannopine synthase gene of T-DNA in maize protoplasts

Multiple ocs-like elements required for efficient transcription of the mannopine synthase gene of T-DNA in maize protoplasts

A 20 nucleotide upstream element is essential for the nopaline synthase (nos) promoter activity

Site-directed mutagenesis of the enhancer region of the 780 gene promoter of T-DNA

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