Multiple transduction pathways regulate the 35S promoter with an ABA responsive element

Chu, Ling-Hui; Jeng, Shih‐Tong

doi:10.1016/s0168-9452(02)00055-9

Cited by 2 publications

(4 citation statements)

References 45 publications

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“…Yet, ABA treatment promoted only a three-fold reduction of PYL6 -CDS mRNA in oxPYL6 as compared to a 54-fold repression in the WT (Figure 2B). Since promoter 35S is not regulated by ABA (Chu and Jeng, 2002), this difference is not due to ABA-mediated transcriptional effect and could, therefore, be a consequence of the inability of mRNA decay regulations to act on the GFP:PYL6 fusion transcript which lacks the 3’UTR region (Fig. 2B).…”

Section: Resultsmentioning

confidence: 99%

Regulation ofPYR/PYL/RCARABA receptors mRNA stability: involvement of miR5628 in decay ofPYL6mRNA

Vieira

Duarte

Barrera-Rojas

et al. 2023

Preprint

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Hormone signaling fine-tuning involves feedback regulatory loops. Abscisic acid (ABA) plays key functions in development and tolerance to abiotic stress. ABA is sensed by the PYR/PYL/RCAR receptors and it also represses their gene expression. Conversely, ABA induces PP2C phosphatases expression, which are negative regulators of the ABA signaling pathway. This feedback regulatory scheme is likely important for the modulation of ABA signal transduction. Here, we provide a new insight into the mechanisms underlying the ABA-induced negative control of PYR/PYL/RCAR expression in Arabidopsis thaliana. The strong and sustained repression of PYR/PYL/RCARs revealed by ABA time course treatment defines the regulation of receptors genes as an important step in resetting the ABA signaling pathway. Transcription inhibition by cordycepin showed that destabilization of PYL1/4/5/6 mRNA is involved in ABA-induced repression of these genes. Furthermore, genetic evidence indicated that decapping may play a role in PYL4/5/6 mRNAs decay. In addition, we provide evidence that the Arabidopsis-specific microRNA5628 (miR5628), which is transiently induced by the ABA core signaling pathway, guides the cleavage of PYL6 transcript in response to ABA. After cleavage, the resulting RISC 5'- and 3'-cleaved fragments of PYL6 mRNA may be degraded by exoribonuclease XRN4. MiR5628 is an evolutionary novelty that may contribute, with decapping and XRN4 activities, to enhance PYL6 mRNA degradation. Thus, control of stability of PYR/PYL/RCAR transcripts is an important step in maintaining homeostasis of ABA signaling.

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Section: Resultsmentioning

confidence: 99%

Regulation ofPYR/PYL/RCARABA receptors mRNA stability: involvement of miR5628 in decay ofPYL6mRNA

Vieira

Duarte

Barrera-Rojas

et al. 2023

Preprint

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“…Plasmid MTC301 containing the 35S promoter and firefly luciferase was kindly provided by Dr. Su-May Yu (Institute of Molecular Biology, Academia Sinica, Taiwan), and was treated as an internal control during electroporation to compare quantitatively the GUS expression from different constructs. The electroporation procedure was reported (Jeng and Yen 2000;Chu and Jeng 2002) and modified as follows. Protoplast isolated from tobacco leaves was adjusted by a dilution solution (0.4 M D-mannitol, 8 mM MES-KOH, and 5 mM CaCl 2 ) to obtain 2 9 10 6 cell/ml.…”

Section: Electroporationmentioning

confidence: 99%

“…The fluorescence of the samples was measured by the excitation wavelength at 365 nm and the emission wavelength at 455 nm, using a Hitachi Spectrofluorometer F2000. A solution of 100 mM 4-methyl umbelliferone (MU) in 0.2 M sodium carbonate was used to calibrate the intensity of fluorescence (Jefferson et al 1987;Jeng and Yen 2000;Chu and Jeng 2002). The concentration of protein in each cell extract was measured by Coomassie Blue Binding method (Sedmak and Grossberg 1977;Spector 1978).…”

Section: Gus Assaymentioning

confidence: 99%

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Effects of the multiple polyadenylation signal AAUAAA on mRNA 3′-end formation and gene expression

Lin

Huang

et al. 2009

Planta

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Polyadenylation (poly(A)) of eukaryotic mRNA is a critical step for gene expression. In plants, poly(A) signals leading to the formation of polyadenosine tails after mRNAs include the far upstream elements, the AAUAAA-like signals, and the mRNA cleavage sites for poly(A). Multiple AAUAAA signals leading to alternative polyadenosine formation have been found in many genes, but the effects of each AAUAAA signal on gene expression remain to be uncovered. A DNA fragment, whose transcript contains two canonical AAUAAA signals from the 3'-untranslation region of endochitinase gene of tobacco (Nicotiana tabacum L. cv. W38), was mutated and constructed into the downstream of beta-glucuronidase (GUS) coding region. Transient expression of GUS gene from these constructs indicated that the distal AAUAAA signal from the stop codon was more important than the proximal one in stimulating gene expression. Also, the sequence rather than the distance between the stop codon and the AAUAAA signal region was critical for gene expression. Transgenic tobaccos with these constructs were also generated, and the position of the polyadenosine tail formation in this region was mapped. Results revealed that both AAUAAA signals were functional, and that polyadenosine tails of most transcripts were directed by the distal AAUAAA signal. Finally, the RNA stabilities of these variants in transgenic plants were measured. RNAs from the variants with the functional distal AAUAAA signal were more stable than those with the functional proximal one only. The possible secondary structure in this poly(A) signal region was predicted and discussed.

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Multiple transduction pathways regulate the 35S promoter with an ABA responsive element

Cited by 2 publications

References 45 publications

Regulation ofPYR/PYL/RCARABA receptors mRNA stability: involvement of miR5628 in decay ofPYL6mRNA

Regulation ofPYR/PYL/RCARABA receptors mRNA stability: involvement of miR5628 in decay ofPYL6mRNA

Effects of the multiple polyadenylation signal AAUAAA on mRNA 3′-end formation and gene expression

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