Potential in vitro and ex vivo targeting of bZIP53 involved in stress response and seed maturation in Arabidopsis thaliana by five designed peptide inhibitors

Jain, Prateek; Shah, Koushik; Rishi, Vikas

doi:10.1016/j.bbapap.2018.09.007

Cited by 6 publications

(10 citation statements)

References 45 publications

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“…A-ZIP53 is designed to specifically target bZIP53 and its known heterodimeric partners and inhibit their DNA binding thus down-regulating associated genes. Earlier in gel shift experiments and transient transfection studies, we have shown the ability of A-ZIP53 in inhibiting DNA binding activities of bZIP53 and its known dimerizing partners i.e., bZIP10 and bZIP25 (Alonso et al 2009;Hartmann et al 2015;Jain et al 2018;Jain et al 2017).…”

Section: Results: A-zip53 Transgene Expression Causes Delayed Growth Phenotype In Arabidopsismentioning

confidence: 89%

“…In a previous study, we have biochemically characterized dominant negative (DN) protein A-ZIP53 and its derivatives. The efficacies of designed proteins were demonstrated in vitro and ex vivo (Jain et al 2018;Jain et al 2017). A-ZIP53 is designed to specifically target bZIP53 and its known heterodimeric partners and inhibit their DNA binding thus down-regulating associated genes.…”

Section: Results: A-zip53 Transgene Expression Causes Delayed Growth Phenotype In Arabidopsismentioning

confidence: 99%

“…bZIP53 binds to DNA as homodimer or as heterodimer with bZIP10, and bZIP25. In earlier we have quantified these interactions using biophysical techniques and gel shift experiments (Jain et al 2018;Jain et al 2017). To investigate the ability of A-ZIP53 in inhibiting the functions of target bZIP53 and its interacting partners, transient transfections were performed using protoplast derived from BY-2 tobacco cell line.…”

Section: Results: A-zip53 Transgene Expression Causes Delayed Growth Phenotype In Arabidopsismentioning

confidence: 99%

“…Earlier, we have functionally validated A-ZIP53 and its derivatives against target bZIP TFs in Arabidopsis (Jain et al 2018;Jain et al 2017). A-ZIP53 has a dimerization domain of bZIP53 and a designed polyglutamic rich acidic extension at its N-terminal.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, we have reported a novel dominant negative (DN) protein approach to regulate the dimeric interaction of target bZIP TFs involved in seed maturation (Jain et al 2018;Jain et al 2017). The efficacy of novel protein A-ZIP53 and its derivatives against wild-type bZIP53 and its heterodimerizing partners were shown in vitro and in vivo (Jain et al 2018;Jain et al 2017). Here, we have generated transgenic Arabidopsis expressing DN protein A-ZIP53 under the constitutive promoter and examined the effects on the growth and development.…”

Section: Introductionmentioning

confidence: 99%

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Synergistic regulation of bZIP53 and dimerizing partners results in abnormal seed phenotype in Arabidopsis: Use of a designed dominant negative protein A-ZIP53

Jain

Rishi

2021

Preprint

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In Arabidopsis, basic leucine zipper (bZIP) family of transcription factors (TFs) are key proteins to regulate the expression of seed maturation (MAT) genes. bZIPs are functionally redundant and their DNA-binding activity is dependent on dimerization partner. The intervention of loss of function mutation is inadequate to understand and regulate the redundant behavior of TFs and one such example is bZIP53, which is known as a key regulator of seed maturation phenomena. Here, to examine the consequences of hindering the function of bZIP53 and its known and unknown heterodimerizing partners, a transgenic Arabidopsis constitutively expressing a novel dominant negative (DN) protein A-ZIP53 was raised. Transgenic plants demonstrated a delayed growth and retarded seed phenotype. The in vivo inhibition of DNA binding of bZIP53, bZIP10, and bZIP25 to the G-box demonstrated the efficacy of A-ZIP53 protein. In first generation, majority of plants failed to survive beyond four weeks suggesting a pleiotropic nature of bZIP53. Plants expressing A-ZIP53 have small flower, shorter siliques, and small-seeded phenotype. RNA seq analysis of the transgenic lines revealed the reduced expression of target genes of bZIP53 and its heterodimerizing partners. Furthermore, immunoprecipitation followed by mass spectrometry (IP-MS/MS) of transgenic plants helped to identify the additional heterodimerizing partners of the A-ZIP53. The interactions were subsequently confirmed with the transient transfection experiments. Unlike other gene knock out technologies, DN protein can inhibit the function of members of the same group of bZIP TFs.

show abstract

Section: Results: A-zip53 Transgene Expression Causes Delayed Growth Phenotype In Arabidopsismentioning

confidence: 89%

Section: Results: A-zip53 Transgene Expression Causes Delayed Growth Phenotype In Arabidopsismentioning

confidence: 99%

Section: Results: A-zip53 Transgene Expression Causes Delayed Growth Phenotype In Arabidopsismentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Synergistic regulation of bZIP53 and dimerizing partners results in abnormal seed phenotype in Arabidopsis: Use of a designed dominant negative protein A-ZIP53

Jain

Rishi

2021

Preprint

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Potential in vitro and ex vivo targeting of bZIP53 involved in stress response and seed maturation in Arabidopsis thaliana by five designed peptide inhibitors

Cited by 6 publications

References 45 publications

Synergistic regulation of bZIP53 and dimerizing partners results in abnormal seed phenotype in Arabidopsis: Use of a designed dominant negative protein A-ZIP53

Synergistic regulation of bZIP53 and dimerizing partners results in abnormal seed phenotype in Arabidopsis: Use of a designed dominant negative protein A-ZIP53

Resistant starch: biosynthesis, regulatory pathways, and engineering via CRISPR system

Salmonella typhimurium detection and ablation using OmpD specific aptamer with non-magnetic and magnetic graphene oxide

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