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1.
Mol Cell ; 81(22): 4591-4604.e8, 2021 11 18.
Artigo em Inglês | MEDLINE | ID: mdl-34592134

RESUMO

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.


Assuntos
ADP-Ribosilação , Proteínas de Arabidopsis/metabolismo , Arabidopsis/imunologia , Proteínas de Ligação a DNA/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Imunidade Vegetal , Ubiquitinação , Dedos de Zinco , ADP Ribose Transferases/metabolismo , Difosfato de Adenosina/química , Arabidopsis/metabolismo , Sistemas CRISPR-Cas , Genes de Plantas , Glicosídeo Hidrolases/metabolismo , Homeostase , Humanos , Hidrólise , Mutação , Plantas Geneticamente Modificadas , Poli Adenosina Difosfato Ribose/metabolismo , Poli(ADP-Ribose) Polimerases/metabolismo , Proteostase , Plântula/metabolismo , Especificidade por Substrato , Tristetraprolina/química , Técnicas do Sistema de Duplo-Híbrido , Ubiquitina/química
2.
J Biol Chem ; 298(9): 102255, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35835222

RESUMO

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.


Assuntos
Alanina , Aminoacil-tRNA Sintetases , Arabidopsis , Proteínas de Plantas , Prolina , Biossíntese de Proteínas , Multimerização Proteica , RNA de Transferência , Alanina/química , Alanina/genética , Aminoacil-tRNA Sintetases/química , Aminoacil-tRNA Sintetases/genética , Arabidopsis/enzimologia , Escherichia coli , Proteínas de Plantas/química , Proteínas de Plantas/genética , Prolina/química , Prolina/genética , Biossíntese de Proteínas/genética , Conformação Proteica em alfa-Hélice , Domínios Proteicos , RNA de Transferência/química
3.
Nucleic Acids Res ; 47(10): 5001-5015, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-30892623

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Proliferação de Células/genética , Fosfatidilinositol 3-Quinases/genética , Raízes de Plantas/genética , Proteínas Repressoras/genética , Fatores de Transcrição/genética , Arabidopsis/citologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Relógios Circadianos , Regulação da Expressão Gênica de Plantas , Meristema/citologia , Meristema/genética , Meristema/metabolismo , Mutação , Fosfatidilinositol 3-Quinases/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/metabolismo , Proteínas Repressoras/metabolismo , Transdução de Sinais/genética , Fatores de Transcrição/metabolismo
4.
BMC Plant Biol ; 18(1): 373, 2018 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-30587139

RESUMO

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.


Assuntos
Brassica rapa/genética , Proteínas de Ligação a DNA/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Dedos de Zinco/genética , Arabidopsis/genética , Brassica rapa/fisiologia , Mapeamento Cromossômico , Sequência Conservada/genética , Proteínas de Ligação a DNA/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/fisiologia , Estudo de Associação Genômica Ampla , Filogenia , Proteínas de Plantas/fisiologia , Alinhamento de Sequência , Estresse Fisiológico , Dedos de Zinco/fisiologia
5.
Plant J ; 78(3): 452-67, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24635033

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/metabolismo , Estabilidade de RNA , RNA de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Elementos Ricos em Adenilato e Uridilato , Motivos de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Sequência de Bases , Sítios de Ligação , Polarização de Fluorescência , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Dados de Sequência Molecular , Mutação , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Fatores de Transcrição/genética , Tristetraprolina/metabolismo , Dedos de Zinco
6.
Plant Cell Physiol ; 55(8): 1367-75, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24850834

RESUMO

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.


Assuntos
Desenvolvimento Vegetal , Proteínas de Plantas/classificação , Plantas/genética , Estresse Fisiológico , Proteínas de Plantas/genética , Plantas/metabolismo , RNA de Plantas/metabolismo , Sequências de Repetição em Tandem , Dedos de Zinco
7.
Front Plant Sci ; 15: 1375478, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38799098

RESUMO

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.
Artigo em Inglês | MEDLINE | ID: mdl-23421766

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Reguladores de Crescimento de Plantas/farmacologia , Sementes/fisiologia , Ácido Abscísico/farmacologia , Arabidopsis/citologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Grânulos Citoplasmáticos , Flores/citologia , Flores/efeitos dos fármacos , Flores/genética , Flores/fisiologia , Genes Reporter , Germinação , Giberelinas/farmacologia , Modelos Biológicos , Mutação , Especificidade de Órgãos , Dormência de Plantas , Folhas de Planta/citologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/fisiologia , Plantas Geneticamente Modificadas , Plântula/citologia , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/fisiologia , Sementes/citologia , Sementes/efeitos dos fármacos , Sementes/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Dedos de Zinco
9.
Plant J ; 65(2): 253-68, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21223390

RESUMO

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.


Assuntos
Ácido Abscísico/metabolismo , Proteínas de Arabidopsis/metabolismo , Arabidopsis/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Giberelinas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Ácido Abscísico/farmacologia , Ácido Abscísico/fisiologia , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Temperatura Baixa , Secas , Expressão Gênica , Germinação , Giberelinas/farmacologia , Giberelinas/fisiologia , Glucose/metabolismo , Hexoquinase/metabolismo , Análise em Microsséries , Reguladores de Crescimento de Plantas/farmacologia , Reguladores de Crescimento de Plantas/fisiologia , Plantas Geneticamente Modificadas , Interferência de RNA , RNA de Plantas/metabolismo , Sementes/crescimento & desenvolvimento , Sementes/fisiologia , Transdução de Sinais/fisiologia , Estresse Fisiológico , Fatores de Transcrição/genética
10.
Plants (Basel) ; 11(4)2022 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-35214845

RESUMO

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.
Artigo em Inglês | MEDLINE | ID: mdl-19897605

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/metabolismo , DNA de Plantas/metabolismo , Transporte Proteico/fisiologia , RNA de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Motivos de Aminoácidos , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Dados de Sequência Molecular , Ligação Proteica , Fatores de Transcrição/genética
12.
Plants (Basel) ; 9(9)2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32872650

RESUMO

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.
Artigo em Inglês | MEDLINE | ID: mdl-31312784

RESUMO

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.
Artigo em Inglês | MEDLINE | ID: mdl-31433992

RESUMO

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.


Assuntos
Arabidopsis/genética , Arabidopsis/imunologia , Imunidade Vegetal/genética , Processamento Pós-Transcricional do RNA/genética , Estabilidade de RNA/genética , Ribonucleoproteínas/metabolismo , Arabidopsis/efeitos dos fármacos , Arabidopsis/microbiologia , Proteínas de Arabidopsis/metabolismo , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Flagelina/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Moléculas com Motivos Associados a Patógenos/metabolismo , Fosforilação/efeitos dos fármacos , Fosfosserina/metabolismo , Imunidade Vegetal/efeitos dos fármacos , Processamento Pós-Transcricional do RNA/efeitos dos fármacos
15.
Plant Sci ; 252: 118-124, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27717446

RESUMO

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.


Assuntos
Motivos de Aminoácidos/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/fisiologia , Domínios e Motivos de Interação entre Proteínas , Estresse Fisiológico , Germinação/genética , Proteínas de Plantas/química , Proteínas de Plantas/genética , Processamento de Proteína Pós-Traducional , RNA Mensageiro/metabolismo , Sementes/genética , Sementes/crescimento & desenvolvimento , Dedos de Zinco
16.
PLoS One ; 11(3): e0151574, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26978070

RESUMO

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.


Assuntos
Ácido Abscísico/fisiologia , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Proteínas de Transporte/metabolismo , Cisteína Proteases/metabolismo , Desidratação/fisiopatologia , Estresse Fisiológico/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/ultraestrutura , Secas , Regulação da Expressão Gênica de Plantas , Genes Reporter , Vetores Genéticos/genética , Imunoprecipitação , Microscopia de Fluorescência , Organelas/fisiologia , Fragmentos de Peptídeos/metabolismo , Mapeamento de Interação de Proteínas , Protoplastos/metabolismo , Protoplastos/ultraestrutura , Processamento Pós-Transcricional do RNA , RNA de Plantas/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Técnicas do Sistema de Duplo-Híbrido , Dedos de Zinco
17.
Plant Sci ; 246: 1-10, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26993231

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Giberelinas/farmacologia , Ácido Abscísico/metabolismo , Arabidopsis/efeitos dos fármacos , Proteínas de Arabidopsis/metabolismo , Flores/efeitos dos fármacos , Flores/genética , Flores/fisiologia , Immunoblotting , Reguladores de Crescimento de Plantas/farmacologia , Plantas Geneticamente Modificadas , Sinais Direcionadores de Proteínas , Transporte Proteico/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Frações Subcelulares/efeitos dos fármacos , Frações Subcelulares/metabolismo
18.
Rice (N Y) ; 9(1): 63, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27888467

RESUMO

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): 647-51, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21795857

RESUMO

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.


Assuntos
Regulação da Expressão Gênica de Plantas , Desenvolvimento Vegetal , Plantas/genética , Estresse Fisiológico/genética , Sequências de Repetição em Tandem/genética , Dedos de Zinco/genética , Sequência de Aminoácidos , Dados de Sequência Molecular , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo
20.
Plant Signal Behav ; 6(5): 719-22, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21455027

RESUMO

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.


Assuntos
Proteínas de Arabidopsis/metabolismo , Proteínas Repressoras/metabolismo , Transativadores/metabolismo , Fatores de Transcrição/metabolismo , Zea mays/genética , Sequência de Aminoácidos , Proteínas de Arabidopsis/química , Dados de Sequência Molecular , Protoplastos/metabolismo , Proteínas Repressoras/química , Saccharomyces cerevisiae/genética , Transativadores/química , Fatores de Transcrição/química , Ativação Transcricional
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