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1.
Mol Cell ; 81(22): 4591-4604.e8, 2021 11 18.
Artículo en Inglés | MEDLINE | ID: mdl-34592134

RESUMEN

Protein ADP-ribosylation is a reversible post-translational modification that transfers ADP-ribose from NAD+ onto acceptor proteins. Poly(ADP-ribosyl)ation (PARylation), catalyzed by poly(ADP-ribose) polymerases (PARPs) and poly(ADP-ribose) glycohydrolases (PARGs), which remove the modification, regulates diverse cellular processes. However, the chemistry and physiological functions of mono(ADP-ribosyl)ation (MARylation) remain elusive. Here, we report that Arabidopsis zinc finger proteins SZF1 and SZF2, key regulators of immune gene expression, are MARylated by the noncanonical ADP-ribosyltransferase SRO2. Immune elicitation promotes MARylation of SZF1/SZF2 via dissociation from PARG1, which has an unconventional activity in hydrolyzing both poly(ADP-ribose) and mono(ADP-ribose) from acceptor proteins. MARylation antagonizes polyubiquitination of SZF1 mediated by the SH3 domain-containing proteins SH3P1/SH3P2, thereby stabilizing SZF1 proteins. Our study uncovers a noncanonical ADP-ribosyltransferase mediating MARylation of immune regulators and underpins the molecular mechanism of maintaining protein homeostasis by the counter-regulation of ADP-ribosylation and polyubiquitination to ensure proper immune responses.


Asunto(s)
ADP-Ribosilación , Proteínas de Arabidopsis/metabolismo , Arabidopsis/inmunología , Proteínas de Unión al ADN/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Inmunidad de la Planta , Ubiquitinación , Dedos de Zinc , ADP Ribosa Transferasas/metabolismo , Adenosina Difosfato/química , Arabidopsis/metabolismo , Sistemas CRISPR-Cas , Genes de Plantas , Glicósido Hidrolasas/metabolismo , Homeostasis , Humanos , Hidrólisis , Mutación , Plantas Modificadas Genéticamente , Poli Adenosina Difosfato Ribosa/metabolismo , Poli(ADP-Ribosa) Polimerasas/metabolismo , Proteostasis , Plantones/metabolismo , Especificidad por Sustrato , Tristetraprolina/química , Técnicas del Sistema de Dos Híbridos , Ubiquitina/química
2.
J Biol Chem ; 298(9): 102255, 2022 09.
Artículo en Inglés | MEDLINE | ID: mdl-35835222

RESUMEN

Faithful translation of the genetic code is critical for the viability of all living organisms. The trans-editing enzyme ProXp-ala prevents Pro to Ala mutations during translation by hydrolyzing misacylated Ala-tRNAPro that has been synthesized by prolyl-tRNA synthetase. Plant ProXp-ala sequences contain a conserved C-terminal domain (CTD) that is absent in other organisms; the origin, structure, and function of this extra domain are unknown. To characterize the plant-specific CTD, we performed bioinformatics and computational analyses that provided a model consistent with a conserved α-helical structure. We also expressed and purified wildtype Arabidopsis thaliana (At) ProXp-ala in Escherichia coli, as well as variants lacking the CTD or containing only the CTD. Circular dichroism spectroscopy confirmed a loss of α-helical signal intensity upon CTD truncation. Size-exclusion chromatography with multiangle laser-light scattering revealed that wildtype At ProXp-ala was primarily dimeric and CTD truncation abolished dimerization in vitro. Furthermore, bimolecular fluorescence complementation assays in At protoplasts support a role for the CTD in homodimerization in vivo. The deacylation rate of Ala-tRNAPro by At ProXp-ala was also significantly reduced in the absence of the CTD, and kinetic assays indicated that the reduction in activity is primarily due to a tRNA binding defect. Overall, these results broaden our understanding of eukaryotic translational fidelity in the plant kingdom. Our study reveals that the plant-specific CTD plays a significant role in substrate binding and canonical editing function. Through its ability to facilitate protein-protein interactions, we propose the CTD may also provide expanded functional potential for trans-editing enzymes in plants.


Asunto(s)
Alanina , Aminoacil-ARNt Sintetasas , Arabidopsis , Proteínas de Plantas , Prolina , Biosíntesis de Proteínas , Multimerización de Proteína , ARN de Transferencia , Alanina/química , Alanina/genética , Aminoacil-ARNt Sintetasas/química , Aminoacil-ARNt Sintetasas/genética , Arabidopsis/enzimología , Escherichia coli , Proteínas de Plantas/química , Proteínas de Plantas/genética , Prolina/química , Prolina/genética , Biosíntesis de Proteínas/genética , Conformación Proteica en Hélice alfa , Dominios Proteicos , ARN de Transferencia/química
3.
Nucleic Acids Res ; 47(10): 5001-5015, 2019 06 04.
Artículo en Inglés | MEDLINE | ID: mdl-30892623

RESUMEN

Circadian clock coordinates numerous plant growth and developmental processes including cell elongation in the hypocotyl, whether or not it modulates cell proliferation is largely unknown. Here we have found that Pseudo Response Regulators (PRRs), essential components of circadian core oscillators, affect root meristem cell proliferation mediated by Target Of Rapamycin (TOR) signaling. The null mutants of PRRs display much reduced sensitivities to sugar-activated TOR signaling. We have subsequently identified Tandem Zinc Finger 1, encoding a processing body localized RNA-binding protein, as a direct target repressed by PRRs in mediating TOR signaling. Multiple lines of biochemical and genetic evidence have demonstrated that TZF1 acts downstream of PRRs to attenuate TOR signaling. Furthermore, TZF1 could directly bind TOR mRNA via its tandem zinc finger motif to affect TOR mRNA stability. Our findings support a notion that PRR-TZF1-TOR molecular axis modulates root meristem cell proliferation by integrating both transcriptional and post-transcriptional regulatory mechanisms.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Proliferación Celular/genética , Fosfatidilinositol 3-Quinasas/genética , Raíces de Plantas/genética , Proteínas Represoras/genética , Factores de Transcripción/genética , Arabidopsis/citología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Relojes Circadianos , Regulación de la Expresión Génica de las Plantas , Meristema/citología , Meristema/genética , Meristema/metabolismo , Mutación , Fosfatidilinositol 3-Quinasas/metabolismo , Raíces de Plantas/citología , Raíces de Plantas/metabolismo , Proteínas Represoras/metabolismo , Transducción de Señal/genética , Factores de Transcripción/metabolismo
4.
BMC Plant Biol ; 18(1): 373, 2018 Dec 27.
Artículo en Inglés | MEDLINE | ID: mdl-30587139

RESUMEN

BACKGROUND: Ubiquitous CCCH nucleic acid-binding motif is found in a wide-variety of organisms. CCCH genes are involved in plant developmental processes and biotic and abiotic stress responses. Brassica rapa is a vital economic crop and classical model plant of polyploidy evolution, but the functions of CCCH genes in B. rapa are unclear. RESULTS: In this study, 103 CCCH genes in B. rapa were identified. A comparative analysis of the chromosomal position, gene structure, domain organization and duplication event between B. rapa and Arabidopsis thaliana were performed. Results showed that CCCH genes could be divided into 18 subfamilies, and segmental duplication might mainly contribute to this family expansion. C-X7/8-C-X5-C3-H was the most commonly found motif, but some novel CCCH motifs were also found, along with some loses of typical CCCH motifs widespread in other plant species. The multifarious gene structures and domain organizations implicated functional diversity of CCCH genes in B. rapa. Evidence also suggested functional redundancy in at least one subfamily due to high conservation between members. Finally, the expression profiles of subfamily-IX genes indicated that they are likely involved in various stress responses. CONCLUSION: This study provides the first genome-wide characterization of the CCCH genes in B. rapa. The results suggest that B. rapa CCCH genes are likely functionally divergent, but mostly involved in plant development and stress response. These results are expected to facilitate future functional characterization of this potential RNA-binding protein family in Brassica crops.


Asunto(s)
Brassica rapa/genética , Proteínas de Unión al ADN/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Dedos de Zinc/genética , Arabidopsis/genética , Brassica rapa/fisiología , Mapeo Cromosómico , Secuencia Conservada/genética , Proteínas de Unión al ADN/fisiología , Regulación de la Expresión Génica de las Plantas/genética , Genes de Plantas/fisiología , Estudio de Asociación del Genoma Completo , Filogenia , Proteínas de Plantas/fisiología , Alineación de Secuencia , Estrés Fisiológico , Dedos de Zinc/fisiología
5.
Plant J ; 78(3): 452-67, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24635033

RESUMEN

The Arabidopsis thaliana tandem zinc finger 1 (AtTZF1) protein is characterized by two tandem-arrayed CCCH-type zinc fingers. We have previously found that AtTZF1 affects hormone-mediated growth, stress and gene expression responses. While much has been learned at the genetic and physiological level, the molecular mechanisms underlying the effects of AtTZF1 on gene expression remain obscure. A human TZF protein, hTTP, is known to bind and trigger the degradation of mRNAs containing AU-rich elements (AREs) at the 3' untranslated regions. However, while the TZF motif of hTTP is characterized by C(X8)C(X5)C(X3)H-(X18)-C(X8)C(X5)C(X3)H, AtTZF1 contains an atypical motif of C(X7)C(X5)C(X3)H-(X16)-C(X5)C(X4)C(X3)H. Moreover, the TZF motif of AtTZF1 is preceded by an arginine-rich (RR) region that is unique to plants. Using fluorescence anisotropy and electrophoretic mobility shift binding assays, we have demonstrated that AtTZF1 binds to RNA molecules with specificity and the interaction is dependent on the presence of zinc. Compared with hTTP, in which TZF is solely responsible for RNA binding, both TZF and RR regions of AtTZF1 are required to achieve high-affinity RNA binding. Moreover, zinc finger integrity is vital for RNA binding. Using a plant protoplast transient expression analysis we have further revealed that AtTZF1 can trigger the decay of ARE-containing mRNAs in vivo. Taken together, our results support the notion that AtTZF1 is involved in RNA turnover.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Estabilidad del ARN , ARN de Planta/metabolismo , Factores de Transcripción/metabolismo , Elementos Ricos en Adenilato y Uridilato , Secuencias de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Secuencia de Bases , Sitios de Unión , Polarización de Fluorescencia , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Datos de Secuencia Molecular , Mutación , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Factores de Transcripción/genética , Tristetraprolina/metabolismo , Dedos de Zinc
6.
Plant Cell Physiol ; 55(8): 1367-75, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24850834

RESUMEN

Cysteine3Histidine (CCCH)-type zinc finger proteins comprise a large family that is well conserved across eukaryotes. Among them, tandem CCCH zinc finger proteins (TZFs) play critical roles in mRNA metabolism in animals and yeast. While there are only three TZF members in humans, a much higher number of TZFs has been found in many plant species. Notably, plant TZFs are over-represented by a class of proteins containing a unique TZF domain preceded by an arginine (R)-rich (RR) motif, hereafter called RR-TZF. Recently, there have been a large number of reports indicating that RR-TZF proteins can localize to processing bodies (P-bodies) and stress granules (SG), two novel cytoplasmic aggregations of messenger ribonucleoprotein complexes (mRNPs), and play critical roles in plant growth, development and stress response, probably via RNA regulation. This review focuses on the classification and most recent development of molecular, cellular and genetic analyses of plant RR-TZF proteins.


Asunto(s)
Desarrollo de la Planta , Proteínas de Plantas/clasificación , Plantas/genética , Estrés Fisiológico , Proteínas de Plantas/genética , Plantas/metabolismo , ARN de Planta/metabolismo , Secuencias Repetidas en Tándem , Dedos de Zinc
7.
Front Plant Sci ; 15: 1375478, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38799098

RESUMEN

Tandem CCCH zinc finger (TZF) proteins play diverse roles in plant growth and stress response. Although as many as 11 TZF proteins have been identified in Arabidopsis, little is known about the mechanism by which TZF proteins select and regulate the target mRNAs. Here, we report that Arabidopsis TZF1 is a bona-fide stress granule protein. Ectopic expression of TZF1 (TZF1 OE), but not an mRNA binding-defective mutant (TZF1H186Y OE), enhances salt stress tolerance in Arabidopsis. RNA-seq analyses of NaCl-treated plants revealed that the down-regulated genes in TZF1 OE plants are enriched for functions in salt and oxidative stress responses. Because many of these down-regulated mRNAs contain AU- and/or U-rich elements (AREs and/or UREs) in their 3'-UTRs, we hypothesized that TZF1-ARE/URE interaction might contribute to the observed gene expression changes. Results from RNA immunoprecipitation-quantitative PCR analysis, gel-shift, and mRNA half-life assays indicate that TZF1 binds and triggers degradation of the autoinhibited Ca2+-ATPase 11 (ACA11) mRNA, which encodes a tonoplast-localized calcium pump that extrudes calcium and dampens signal transduction pathways necessary for salt stress tolerance. Furthermore, this salt stress-tolerance phenotype was recapitulated in aca11 null mutants. Collectively, our findings demonstrate that TZF1 binds and initiates degradation of specific mRNAs to enhance salt stress tolerance.

8.
Plant Cell Environ ; 36(8): 1507-19, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23421766

RESUMEN

Tandem CCCH zinc finger proteins (TZFs) are post-transcriptional regulators of gene expression in animals and yeast. Genetic studies indicate that plant TZFs are involved in hormone-mediated developmental and environmental responses. We have demonstrated previously that Arabidopsis AtTZF1 can localize to processing bodies (PBs) and stress granules (SGs), and affects abscisic acid (ABA)- and gibberellic acid (GA)-mediated growth, stress and gene expression responses. Here we show that AtTZF4, 5 and 6 are specifically expressed in seeds. Consistent with the observation that their expression levels decline during seed imbibition, AtTZF4, 5 and 6 are up-regulated by ABA and down-regulated by GA. Mutant analyses indicate that AtTZF4, 5 and 6 act as positive regulators for ABA- and negative regulators for light- and GA-mediated seed germination responses. Results of gene expression analysis indicate that AtTZF4, 5 and 6 affect seed germination by controlling genes critical for ABA and GA response. Furthermore, AtTZF4, 5 and 6 can co-localize with both PB and SG markers in Arabidopsis cells. Specifically, AtTZF6 can be assembled into PBs and SGs in embryos with the induction of stress hormone methyl jasmonate under the control of native AtTZF6 promoter.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiología , Regulación de la Expresión Génica de las Plantas , Reguladores del Crecimiento de las Plantas/farmacología , Semillas/fisiología , Ácido Abscísico/farmacología , Arabidopsis/citología , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Gránulos Citoplasmáticos , Flores/citología , Flores/efectos de los fármacos , Flores/genética , Flores/fisiología , Genes Reporteros , Germinación , Giberelinas/farmacología , Modelos Biológicos , Mutación , Especificidad de Órganos , Latencia en las Plantas , Hojas de la Planta/citología , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Plantas Modificadas Genéticamente , Plantones/citología , Plantones/efectos de los fármacos , Plantones/genética , Plantones/fisiología , Semillas/citología , Semillas/efectos de los fármacos , Semillas/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Dedos de Zinc
9.
Plant J ; 65(2): 253-68, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21223390

RESUMEN

Tandem zinc finger (TZF) proteins are characterized by two zinc-binding CCCH motifs arranged in tandem. Human TZFs such as tristetraproline (TTP) bind to and trigger the degradation of mRNAs encoding cytokines and various regulators. Although the molecular functions of plant TZFs are unknown, recent genetic studies have revealed roles in hormone-mediated growth and environmental responses, as well as in the regulation of gene expression. Here we show that expression of AtTZF1 (AtCTH/AtC3H23) mRNA is repressed by a hexokinase-dependent sugar signaling pathway. However, AtTZF1 acts as a positive regulator of ABA/sugar responses and a negative regulator of GA responses, at least in part by modulating gene expression. RNAi of AtTZF1-3 caused early germination and slightly stress-sensitive phenotypes, whereas plants over-expressing AtTZF1 were compact, late flowering and stress-tolerant. The developmental phenotypes of plants over-expressing AtTZF1 were only partially rescued by exogenous application of GA, implying a reduction in the GA response or defects in other mechanisms. Likewise, the enhanced cold and drought tolerance of plants over-expressing AtTZF1 were not associated with increased ABA accumulation, suggesting that it is mainly ABA responses that are affected. Consistent with this notion, microarray analysis showed that over-expression of AtTZF1 mimics the effects of ABA or GA deficiency on gene expression. Notably, a gene network centered on a GA-inducible and ABA/sugar-repressible putative peptide hormone encoded by GASA6 was severely repressed by AtTZF1 over-expression. Hence AtTZF1 may serve as a regulator connecting sugar, ABA, GA and peptide hormone responses.


Asunto(s)
Ácido Abscísico/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/crecimiento & desarrollo , Regulación de la Expresión Génica de las Plantas , Giberelinas/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Factores de Transcripción/metabolismo , Ácido Abscísico/farmacología , Ácido Abscísico/fisiología , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Frío , Sequías , Expresión Génica , Germinación , Giberelinas/farmacología , Giberelinas/fisiología , Glucosa/metabolismo , Hexoquinasa/metabolismo , Análisis por Micromatrices , Reguladores del Crecimiento de las Plantas/farmacología , Reguladores del Crecimiento de las Plantas/fisiología , Plantas Modificadas Genéticamente , Interferencia de ARN , ARN de Planta/metabolismo , Semillas/crecimiento & desarrollo , Semillas/fisiología , Transducción de Señal/fisiología , Estrés Fisiológico , Factores de Transcripción/genética
10.
Plants (Basel) ; 11(4)2022 Feb 14.
Artículo en Inglés | MEDLINE | ID: mdl-35214845

RESUMEN

Leaves are essential plant organs with numerous variations in shape and size. The leaf size is generally smaller in plants that thrive in areas of higher elevation and lower annual mean temperature. The Qinghai-Tibetan Plateau is situated at an altitude of >4000 m with relatively low annual average temperatures. Most plant species found on the Qinghai-Tibetan Plateau have small leaves, with Rheum tanguticum Maxim. ex Balf. being an exception. Here, we show that the large leaves of R. tanguticum with a unique three-dimensional (3D) shape are potentially an ideal solution for thermoregulation with little energy consumption. With the increase in age, the shape of R. tanguticum leaves changed from a small oval plane to a large palmatipartite 3D shape. Therefore, R. tanguticum is a highly heteroblastic species. The leaf shape change during the transition from the juvenile to the adult phase of the development in R. tanguticum is a striking example of the manifestation of plant phenotypic plasticity. The temperature variation in different parts of the leaf was a distinct character of leaves of over-5-year-old plants. The temperature of single-plane leaves under strong solar radiation could accumulate heat rapidly and resulted in temperatures much higher than the ambient temperature. However, leaves of over-5-year-old plants could lower leaf temperature by avoiding direct exposure to solar radiation and promoting local airflow to prevent serious tissue damage by sunburn. Furthermore, the net photosynthesis rate was correlated with the heterogeneity of the leaf surface temperature. Our results demonstrate that the robust 3D shape of the leaf is a strategy that R. tanguticum has developed evolutionarily to adapt to the strong solar radiation and low temperature on the Qinghai-Tibetan Plateau.

11.
Plant Physiol ; 152(1): 151-65, 2010 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-19897605

RESUMEN

Processing bodies (PBs) are specialized cytoplasmic foci where mRNA turnover and translational repression can take place. Stress granules are related cytoplasmic foci. The CCCH tandem zinc finger proteins (TZFs) play pivotal roles in gene expression, cell fate specification, and various developmental processes. Human TZF binds AU-rich elements at the 3' untranslated region and recruits decapping, deadenylation, and exonucleolytic enzymes to PBs for RNA turnover. Recent genetic studies indicate that plant TZFs are involved in gene regulation and hormone-mediated environmental responses. It is unknown if plant TZFs can bind RNA and be localized to PBs or stress granules. The Arabidopsis (Arabidopsis thaliana) AtTZF1/AtCTH/AtC3H23 was identified as a sugar-sensitive gene in a previous microarray study. It is characterized by a TZF motif that is distinct from the human TZF. Higher plants such as Arabidopsis and rice (Oryza sativa) each have a gene family containing this unique TZF motif. Here, we show that AtTZF1 can traffic between the nucleus and cytoplasmic foci. AtTZF1 colocalizes with markers of PBs, and the morphology of these cytoplasmic foci resembles that of mammalian PBs and stress granules. AtTZF1-associated cytoplasmic foci are dynamic and tissue specific. They can be induced by dark and wound stresses and are preferentially present in actively growing tissues and stomatal precursor cells. Since AtTZF1 can bind both DNA and RNA in vitro, it raises the possibility that AtTZF1 might be involved in DNA and/or RNA regulation.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , ADN de Plantas/metabolismo , Transporte de Proteínas/fisiología , ARN de Planta/metabolismo , Factores de Transcripción/metabolismo , Secuencias de Aminoácidos , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Regulación de la Expresión Génica de las Plantas/fisiología , Datos de Secuencia Molecular , Unión Proteica , Factores de Transcripción/genética
12.
Plants (Basel) ; 9(9)2020 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-32872650

RESUMEN

RNA granules, such as stress granules and processing bodies, can balance the storage, degradation, and translation of mRNAs in diverse eukaryotic organisms. The sessile nature of plants demands highly versatile strategies to respond to environmental fluctuations. In this review, we discuss recent findings of the dynamics and functions of these RNA granules in plants undergoing developmental reprogramming or responding to environmental stresses. Special foci include the dynamic assembly, disassembly, and regulatory roles of these RNA granules in determining the fate of mRNAs.

13.
Plant Direct ; 3(7): e00142, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31312784

RESUMEN

Improving yield by increasing the size of produce is an important selection criterion during the domestication of fruit and vegetable crops. Genes controlling meristem organization and organ formation work in concert to regulate the size of reproductive organs. In tomato, lc and fas control locule number, which often leads to enlarged fruits compared to the wild progenitors. LC is encoded by the tomato ortholog of WUSCHEL (WUS), whereas FAS is encoded by the tomato ortholog of CLAVATA3 (CLV3). The critical role of the WUS-CLV3 feedback loop in meristem organization has been demonstrated in several plant species. We show that mutant alleles for both loci in tomato led to an expansion of the SlWUS expression domain in young floral buds 2-3 days after initiation. Single and double mutant alleles of lc and fas maintain higher SlWUS expression during the development of the carpel primordia in the floral bud. This augmentation and altered spatial expression of SlWUS provided a mechanistic basis for the formation of multilocular and large fruits. Our results indicated that lc and fas are gain-of-function and partially loss-of-function alleles, respectively, while both mutations positively affect the size of tomato floral meristems. In addition, expression profiling showed that lc and fas affected the expression of several genes in biological processes including those involved in meristem/flower development, patterning, microtubule binding activity, and sterol biosynthesis. Several differentially expressed genes co-expressed with SlWUS have been identified, and they are enriched for functions in meristem regulation. Our results provide new insights into the transcriptional regulation of genes that modulate meristem maintenance and floral organ determinacy in tomato.

14.
Cell Rep ; 28(8): 2194-2205.e6, 2019 08 20.
Artículo en Inglés | MEDLINE | ID: mdl-31433992

RESUMEN

Proper transcriptome reprogramming is critical for hosts to launch an effective defense response upon pathogen attack. How immune-related genes are regulated at the posttranscriptional level remains elusive. We demonstrate here that P-bodies, the non-membranous cytoplasmic ribonucleoprotein foci related to 5'-to-3' mRNA decay, are dynamically modulated in plant immunity triggered by microbe-associated molecular patterns (MAMPs). The DCP1-DCP2 mRNA decapping complex, a hallmark of P-bodies, positively regulates plant MAMP-triggered responses and immunity against pathogenic bacteria. MAMP-activated MAP kinases directly phosphorylate DCP1 at the serine237 residue, which further stimulates its interaction with XRN4, an exonuclease executing 5'-to-3' degradation of decapped mRNA. Consequently, MAMP treatment potentiates DCP1-dependent mRNA decay on a specific group of MAMP-downregulated genes. Thus, the conserved 5'-to-3' mRNA decay elicited by the MAMP-activated MAP kinase cascade is an integral part of plant immunity. This mechanism ensures a rapid posttranscriptional downregulation of certain immune-related genes that may otherwise negatively impact immunity.


Asunto(s)
Arabidopsis/genética , Arabidopsis/inmunología , Inmunidad de la Planta/genética , Procesamiento Postranscripcional del ARN/genética , Estabilidad del ARN/genética , Ribonucleoproteínas/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/microbiología , Proteínas de Arabidopsis/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Flagelina/farmacología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Moléculas de Patrón Molecular Asociado a Patógenos/metabolismo , Fosforilación/efectos de los fármacos , Fosfoserina/metabolismo , Inmunidad de la Planta/efectos de los fármacos , Procesamiento Postranscripcional del ARN/efectos de los fármacos
15.
Plant Sci ; 252: 118-124, 2016 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-27717446

RESUMEN

Tandem CCCH zinc finger (TZF) proteins are evolutionarily conserved regulators of gene expression at the post-transcriptional level. TZFs target AU-rich RNA elements at 3' un-translated region and recruit catabolic machineries to trigger mRNA degradation. The plant TZF families are over-represented by a class of proteins with a unique TZF domain preceded by an arginine-rich motif (RR-TZF). RR-TZF proteins are mainly involved in hormone- and environmental cues-mediated plant growth and stress responses. Numerous reports have suggested that RR-TZF proteins control seed germination, plant size, flowering time, and biotic and abiotic stress responses via regulation of gene expression. Despite growing genetic evidence, the underlying molecular mechanisms are elusive. This review outlines the highly conserved roles of plant RR-TZFs in various stress responses and the potential involvements of RR-TZF proteins in transcriptional and post-transcriptional regulation of gene expression. The dynamic subcellular localization of RR-TZF proteins, implication of predominant protein-protein interactions between RR-TZF proteins and stress response mediators and future directions of this research field are also discussed.


Asunto(s)
Secuencias de Aminoácidos/fisiología , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/fisiología , Dominios y Motivos de Interacción de Proteínas , Estrés Fisiológico , Germinación/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Procesamiento Proteico-Postraduccional , ARN Mensajero/metabolismo , Semillas/genética , Semillas/crecimiento & desarrollo , Dedos de Zinc
16.
PLoS One ; 11(3): e0151574, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26978070

RESUMEN

Although multiple lines of evidence have indicated that Arabidopsis thaliana Tandem CCCH Zinc Finger proteins, AtTZF4, 5 and 6 are involved in ABA, GA and phytochrome mediated seed germination responses, the interacting proteins involved in these processes are unknown. Using yeast two-hybrid screens, we have identified 35 putative AtTZF5 interacting protein partners. Among them, Mediator of ABA-Regulated Dormancy 1 (MARD1) is highly expressed in seeds and involved in ABA signal transduction, while Responsive to Dehydration 21A (RD21A) is a well-documented stress responsive protein. Co-immunoprecipitation (Co-IP) and bimolecular fluorescence complementation (BiFC) assays were used to confirm that AtTZF5 can interact with MARD1 and RD21A in plant cells, and the interaction is mediated through TZF motif. In addition, AtTZF4 and 6 could also interact with MARD1 and RD21A in Y-2-H and BiFC assay, respectively. The protein-protein interactions apparently take place in processing bodies (PBs) and stress granules (SGs), because AtTZF5, MARD1 and RD21A could interact and co-localize with each other and they all can co-localize with the same PB and SG markers in plant cells.


Asunto(s)
Ácido Abscísico/fisiología , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiología , Proteínas Portadoras/metabolismo , Proteasas de Cisteína/metabolismo , Deshidratación/fisiopatología , Estrés Fisiológico/fisiología , Factores de Transcripción/metabolismo , Arabidopsis/ultraestructura , Sequías , Regulación de la Expresión Génica de las Plantas , Genes Reporteros , Vectores Genéticos/genética , Inmunoprecipitación , Microscopía Fluorescente , Orgánulos/fisiología , Fragmentos de Péptidos/metabolismo , Mapeo de Interacción de Proteínas , Protoplastos/metabolismo , Protoplastos/ultraestructura , Procesamiento Postranscripcional del ARN , ARN de Planta/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Técnicas del Sistema de Dos Híbridos , Dedos de Zinc
17.
Plant Sci ; 246: 1-10, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-26993231

RESUMEN

GA and ABA play antagonistic roles in numerous cellular processes essential for growth, development, and stress responses. GASA4 and GASA6 belong to a family of GA-Stimulated transcripts in Arabidopsis, known as GA-inducible and ABA-repressible. We have found that GASA4 and GASA6 expression is likely mediated through a repressor of GA responses, GA INSENSITIVE (GAI) protein. Moreover, GASA4 and GASA6 are in general up regulated by growth hormones (auxin, BR, cytokinin, and GA) and down regulated by stress hormones (ABA, JA, and SA), indicating a role of GASA4 and GASA6 in hormone crosstalk. Genetic analyses show that suppression of both GASA4 and GASA6 causes late flowering, while over-expression of GASA6 causes early flowering in Arabidopsis. GASA family members encode small polypeptides sharing common structural features: an N-terminal signal peptide, a highly divergent intermediate region, and a conserved C-terminal domain containing 12 conserved cysteines. Despite the presence of a signal peptide, it has not been determined whether or not GASA4 and GASA6 can be processed in vivo. By using imaging and immunological analyses, we show that the N-terminal signal peptide is cleaved as predicted, and the cleavage is important for proper sub-cellular localization of GASA4 and GASA6.


Asunto(s)
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Giberelinas/farmacología , Ácido Abscísico/metabolismo , Arabidopsis/efectos de los fármacos , Proteínas de Arabidopsis/metabolismo , Flores/efectos de los fármacos , Flores/genética , Flores/fisiología , Immunoblotting , Reguladores del Crecimiento de las Plantas/farmacología , Plantas Modificadas Genéticamente , Señales de Clasificación de Proteína , Transporte de Proteínas/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Fracciones Subcelulares/efectos de los fármacos , Fracciones Subcelulares/metabolismo
18.
Rice (N Y) ; 9(1): 63, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27888467

RESUMEN

BACKGROUND: Plant WRKY transcription factors play pivotal roles in diverse biological processes but most notably in plant defense response to pathogens. Sheath blight represents one of the predominant diseases in rice. However, our knowledge about the functions of WRKY proteins in rice defense against sheath blight is rather limited. RESULTS: Here we demonstrate that the expression of Oryza sativa WRKY80 gene (OsWRKY80) is rapidly and strongly induced upon infection of Rhizoctonia solani, the causal agent of rice sheath blight disease. OsWRKY80 expression is also induced by exogenous jasmonic acid (JA) and ethylene (ET), but not by salicylic acid (SA). OsWRKY80-GFP is localized in the nuclei of onion epidermal cells in a transient expression assay. Consistently, OsWRKY80 exhibits transcriptional activation activity in a GAL4 assay in yeast cells. Overexpression of OsWRKY80 in rice plants significantly enhanced disease resistance to R. solani, concomitant with elevated expression of OsWRKY4, another positive regulator in rice defense against R. solani. Suppression of OsWRKY80 by RNA interference (RNAi), on the other hand, compromised disease resistance to R. solani. Results of yeast one-hybrid assay and transient expression assay in tobacco cells have revealed that OsWRKY80 specifically binds to the promoter regions of OsWRKY4, which contain W-box (TTGAC[C/T]) or W-box like (TGAC[C/T]) cis-elements. CONCLUSIONS: We propose that OsWRKY80 functions upstream of OsWRKY4 as an important positive regulatory circuit that is implicated in rice defense response to sheath blight pathogen R. solani.

19.
Plant Signal Behav ; 6(5): 719-22, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21455027

RESUMEN

In animals, Tandem CCCH Zinc Finger (TZF) proteins can affect gene expression at both transcriptional and post-transcriptional levels. In Arabidopsis thaliana, AtTZF1 is a member of the TZF family characterized by a plant-unique tandem zinc finger motif. AtTZF1 can bind both DNA and RNA in vitro, and it can traffic between the nucleus and cytoplasmic foci. However, no in vivo DNA/RNA targets have been identified so far, and little is known about the molecular mechanisms underlying AtTZF1's profound effects on plant growth, development, and stress responses. In order to determine whether AtTZF1 can function as a transcription factor, transactivation assays were conducted. Results indicated that AtTZF1 fusion proteins could not exert obvious transcriptional activity in a maize protoplast transient expression system. However, this conclusion might be biased due to poor nuclear localization of AtTZF1 fusion proteins in the assay system.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Proteínas Represoras/metabolismo , Transactivadores/metabolismo , Factores de Transcripción/metabolismo , Zea mays/genética , Secuencia de Aminoácidos , Proteínas de Arabidopsis/química , Datos de Secuencia Molecular , Protoplastos/metabolismo , Proteínas Represoras/química , Saccharomyces cerevisiae/genética , Transactivadores/química , Factores de Transcripción/química , Activación Transcripcional
20.
Plant Signal Behav ; 6(5): 647-51, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21795857

RESUMEN

In animals, Tandem CCCH Zinc Finger (TZF) proteins control a variety of cellular processes via regulation of gene expression at transcriptional and post-transcriptional levels. Plant-unique TZF proteins can also affect many aspects of plant growth, development, and stress responses. However, the molecular mechanisms underlying plant TZF function are unknown. The purpose of this short review is to provide an overview of genetic and molecular analyses of plant TZFs, and to speculate on their possible molecular functions.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Desarrollo de la Planta , Plantas/genética , Estrés Fisiológico/genética , Secuencias Repetidas en Tándem/genética , Dedos de Zinc/genética , Secuencia de Aminoácidos , Datos de Secuencia Molecular , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo
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