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1.
Annu Rev Biochem ; 92: 227-245, 2023 06 20.
Artículo en Inglés | MEDLINE | ID: mdl-37001134

RESUMEN

Messenger RNA (mRNA) stability and translational efficiency are two crucial aspects of the post-transcriptional process that profoundly impact protein production in a cell. While it is widely known that ribosomes produce proteins, studies during the past decade have surprisingly revealed that ribosomes also control mRNA stability in a codon-dependent manner, a process referred to as codon optimality. Therefore, codons, the three-nucleotide words read by the ribosome, have a potent effect on mRNA stability and provide cis-regulatory information that extends beyond the amino acids they encode. While the codon optimality molecular mechanism is still unclear, the translation elongation rate appears to trigger mRNA decay. Thus, transfer RNAs emerge as potential master gene regulators affecting mRNA stability. Furthermore, while few factors related to codon optimality have been identified in yeast, the orthologous genes in vertebrates do not necessary share the same functions. Here, we discuss codon optimality findings and gene regulation layers related to codon composition in different eukaryotic species.


Asunto(s)
Biosíntesis de Proteínas , Proteínas , Animales , ARN Mensajero/metabolismo , Codón/genética , Proteínas/genética , Estabilidad del ARN , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo
2.
Nucleic Acids Res ; 2024 Jun 25.
Artículo en Inglés | MEDLINE | ID: mdl-38917323

RESUMEN

Zika virus (ZIKV) is an emerging mosquito-borne flavivirus that causes severe outbreaks in human populations. ZIKV infection leads to the accumulation of small non-coding viral RNAs (known as sfRNAs) that are crucial for evasion of antiviral responses and for viral pathogenesis. However, the mechanistic understanding of how sfRNAs function remains incomplete. Here, we use recombinant ZIKVs and ribosome profiling of infected human cells to show that sfRNAs block translation of antiviral genes. Mechanistically, we demonstrate that specific RNA structures present in sfRNAs trigger PKR activation, which instead of limiting viral replication, enhances viral particle production. Although ZIKV infection induces mRNA expression of antiviral genes, translation efficiency of type I interferon and interferon stimulated genes were significantly downregulated by PKR activation. Our results reveal a novel viral adaptation mechanism mediated by sfRNAs, where ZIKV increases its fitness by repurposing the antiviral role of PKR into a proviral factor.

3.
Mol Cell ; 65(3): 375-377, 2017 Feb 02.
Artículo en Inglés | MEDLINE | ID: mdl-28157501

RESUMEN

In this issue, Aeschimann et al. (2017) demonstrate that, depending on the target location site (5'UTR or 3'UTR), LIN41 triggers repression of translation or mRNA decay, suggesting that one factor may use two independent pathways of post-transcriptional gene regulation.


Asunto(s)
Biosíntesis de Proteínas , ARN Mensajero/metabolismo , Factores de Transcripción/metabolismo , Regiones no Traducidas 3' , Regiones no Traducidas 5' , Animales , Regulación de la Expresión Génica , Humanos , Estabilidad del ARN , ARN Mensajero/química
4.
EMBO J ; 39(17): e104763, 2020 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-32744758

RESUMEN

In addition to canonical open reading frames (ORFs), thousands of translated small ORFs (containing less than 100 codons) have been identified in untranslated mRNA regions (UTRs) across eukaryotes. Small ORFs in 5' UTRs (upstream (u)ORFs) often repress translation of the canonical ORF within the same mRNA. However, the function of translated small ORFs in the 3' UTRs (downstream (d)ORFs) is unknown. Contrary to uORFs, we find that translation of dORFs enhances translation of their corresponding canonical ORFs. This translation stimulatory effect of dORFs depends on the number of dORFs, but not the length or peptide they encode. We propose that dORFs represent a new, strong, and universal translation regulatory mechanism in vertebrates.


Asunto(s)
Codón , Sistemas de Lectura Abierta , Biosíntesis de Proteínas , Proteínas de Pez Cebra , Pez Cebra , Animales , Codón/genética , Codón/metabolismo , Pez Cebra/genética , Pez Cebra/metabolismo , Proteínas de Pez Cebra/biosíntesis , Proteínas de Pez Cebra/genética
5.
EMBO J ; 35(7): 706-23, 2016 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-26896445

RESUMEN

Regulation of gene expression is fundamental in establishing cellular diversity and a target of natural selection. Untranslated mRNA regions (UTRs) are key mediators of post-transcriptional regulation. Previous studies have predicted thousands of ORFs in 5'UTRs, the vast majority of which have unknown function. Here, we present a systematic analysis of the translation and function of upstream open reading frames (uORFs) across vertebrates. Using high-resolution ribosome footprinting, we find that (i)uORFs are prevalent within vertebrate transcriptomes, (ii) the majority show signatures of active translation, and (iii)uORFs act as potent regulators of translation and RNA levels, with a similar magnitude to miRNAs. Reporter experiments reveal clear repression of downstream translation by uORFs/oORFs. uORF number, intercistronic distance, overlap with the CDS, and initiation context most strongly influence translation. Evolution has targeted these features to favor uORFs amenable to regulation over constitutively repressive uORFs/oORFs. Finally, we observe that the regulatory potential of uORFs on individual genes is conserved across species. These results provide insight into the regulatory code within mRNA leader sequences and their capacity to modulate translation across vertebrates.


Asunto(s)
Sistemas de Lectura Abierta , Biosíntesis de Proteínas , Proteínas Represoras/metabolismo , Vertebrados/genética , Animales , Transcripción Genética
6.
EMBO J ; 35(19): 2087-2103, 2016 10 04.
Artículo en Inglés | MEDLINE | ID: mdl-27436874

RESUMEN

Cellular transitions require dramatic changes in gene expression that are supported by regulated mRNA decay and new transcription. The maternal-to-zygotic transition is a conserved developmental progression during which thousands of maternal mRNAs are cleared by post-transcriptional mechanisms. Although some maternal mRNAs are targeted for degradation by microRNAs, this pathway does not fully explain mRNA clearance. We investigated how codon identity and translation affect mRNA stability during development and homeostasis. We show that the codon triplet contains translation-dependent regulatory information that influences transcript decay. Codon composition shapes maternal mRNA clearance during the maternal-to-zygotic transition in zebrafish, Xenopus, mouse, and Drosophila, and gene expression during homeostasis across human tissues. Some synonymous codons show consistent stabilizing or destabilizing effects, suggesting that amino acid composition influences mRNA stability. Codon composition affects both polyadenylation status and translation efficiency. Thus, the ribosome interprets two codes within the mRNA: the genetic code which specifies the amino acid sequence and a conserved "codon optimality code" that shapes mRNA stability and translation efficiency across vertebrates.


Asunto(s)
Codón , Regulación de la Expresión Génica , Biosíntesis de Proteínas , Estabilidad del ARN , ARN Mensajero/genética , Cigoto/crecimiento & desarrollo , Animales , Drosophila , Humanos , Ratones , Ribosomas/metabolismo , Xenopus , Pez Cebra
7.
Nature ; 503(7476): 360-4, 2013 Nov 21.
Artículo en Inglés | MEDLINE | ID: mdl-24056933

RESUMEN

After fertilization, maternal factors direct development and trigger zygotic genome activation (ZGA) at the maternal-to-zygotic transition (MZT). In zebrafish, ZGA is required for gastrulation and clearance of maternal messenger RNAs, which is in part regulated by the conserved microRNA miR-430. However, the factors that activate the zygotic program in vertebrates are unknown. Here we show that Nanog, Pou5f1 (also called Oct4) and SoxB1 regulate zygotic gene activation in zebrafish. We identified several hundred genes directly activated by maternal factors, constituting the first wave of zygotic transcription. Ribosome profiling revealed that nanog, sox19b and pou5f1 are the most highly translated transcription factors pre-MZT. Combined loss of these factors resulted in developmental arrest before gastrulation and a failure to activate >75% of zygotic genes, including miR-430. Our results demonstrate that maternal Nanog, Pou5f1 and SoxB1 are required to initiate the zygotic developmental program and induce clearance of the maternal program by activating miR-430 expression.


Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/metabolismo , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , Factores de Transcripción SOXB1/metabolismo , Proteínas de Pez Cebra/metabolismo , Pez Cebra/embriología , Pez Cebra/genética , Cigoto/metabolismo , Animales , Reprogramación Celular/genética , Desarrollo Embrionario/genética , Femenino , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica/genética , MicroARNs/genética , Madres , Proteína Homeótica Nanog , Células Madre Pluripotentes/metabolismo , Ribosomas/genética , Transcriptoma/genética
8.
EMBO J ; 33(9): 981-93, 2014 May 02.
Artículo en Inglés | MEDLINE | ID: mdl-24705786

RESUMEN

Identification of the coding elements in the genome is a fundamental step to understanding the building blocks of living systems. Short peptides (< 100 aa) have emerged as important regulators of development and physiology, but their identification has been limited by their size. We have leveraged the periodicity of ribosome movement on the mRNA to define actively translated ORFs by ribosome footprinting. This approach identifies several hundred translated small ORFs in zebrafish and human. Computational prediction of small ORFs from codon conservation patterns corroborates and extends these findings and identifies conserved sequences in zebrafish and human, suggesting functional peptide products (micropeptides). These results identify micropeptide-encoding genes in vertebrates, providing an entry point to define their function in vivo.


Asunto(s)
Secuencia Conservada , Evolución Molecular , Sistemas de Lectura Abierta/genética , ARN Mensajero/genética , Ribosomas/metabolismo , Pez Cebra/genética , Animales , Secuencia de Bases , Biología Computacional , Perfilación de la Expresión Génica , Humanos , Datos de Secuencia Molecular , Ensayos de Protección de Nucleasas , Oligopéptidos/genética , ARN Mensajero/metabolismo , Análisis de Secuencia de ARN/métodos , Pez Cebra/embriología
10.
Genome Biol ; 25(1): 74, 2024 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-38504288

RESUMEN

BACKGROUND: Early embryonic developmental programs are guided by the coordinated interplay between maternally inherited and zygotically manufactured RNAs and proteins. Although these processes happen concomitantly and affecting gene function during this period is bound to affect both pools of mRNAs, it has been challenging to study their expression dynamics separately. RESULTS: By employing SLAM-seq, a nascent mRNA labeling transcriptomic approach, in a developmental time series we observe that over half of the early zebrafish embryo transcriptome consists of maternal-zygotic genes, emphasizing their pivotal role in early embryogenesis. We provide an hourly resolution of de novo transcriptional activation events and follow nascent mRNA trajectories, finding that most de novo transcriptional events are stable throughout this period. Additionally, by blocking microRNA-430 function, a key post transcriptional regulator during zebrafish embryogenesis, we directly show that it destabilizes hundreds of de novo transcribed mRNAs from pure zygotic as well as maternal-zygotic genes. This unveils a novel miR-430 function during embryogenesis, fine-tuning zygotic gene expression. CONCLUSION: These insights into zebrafish early embryo transcriptome dynamics emphasize the significance of post-transcriptional regulators in zygotic genome activation. The findings pave the way for future investigations into the coordinated interplay between transcriptional and post-transcriptional landscapes required for the establishment of animal cell identities and functions.


Asunto(s)
MicroARNs , Pez Cebra , Animales , Pez Cebra/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Cigoto/metabolismo , Desarrollo Embrionario/genética , MicroARNs/genética , MicroARNs/metabolismo , Regulación del Desarrollo de la Expresión Génica
11.
bioRxiv ; 2024 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-38826327

RESUMEN

The Maternal-to-Zygotic transition (MZT) is a reprograming process encompassing zygotic genome activation (ZGA) and the clearance of maternally-provided mRNAs. While some factors regulating MZT have been identified, there are thousands of maternal RNAs whose function has not been ascribed yet. Here, we have performed a proof-of-principle CRISPR-RfxCas13d maternal screening targeting mRNAs encoding protein kinases and phosphatases in zebrafish and identified Bckdk as a novel post-translational regulator of MZT. Bckdk mRNA knockdown caused epiboly defects, ZGA deregulation, H3K27ac reduction and a partial impairment of miR-430 processing. Phospho-proteomic analysis revealed that Phf10/Baf45a, a chromatin remodeling factor, is less phosphorylated upon Bckdk depletion. Further, phf10 mRNA knockdown also altered ZGA and Phf10 constitutively phosphorylated rescued the developmental defects observed after bckdk mRNA depletion. Altogether, our results demonstrate the competence of CRISPR-RfxCas13d screenings to uncover new regulators of early vertebrate development and shed light on the post-translational control of MZT mediated by protein phosphorylation.

12.
STAR Protoc ; 3(1): 101058, 2022 03 18.
Artículo en Inglés | MEDLINE | ID: mdl-35005640

RESUMEN

CRISPR-Cas systems have been used to induce DNA mutagenesis for gene function discovery. However, the development of tools to eliminate RNAs provides complementary and unique approaches to disrupt gene expression. Here, we present a workflow to perform specific, efficient, and cost-effective mRNA knockdown in zebrafish embryos using our in vivo optimized CRISPR-RfxCas13d (CasRx) system. Although the described protocol focuses on mRNA knockdown in zebrafish embryos, it can also be applied to other vertebrates. For complete details on the use and execution of this protocol, please refer to Kushawah et al. (2020).


Asunto(s)
ARN Guía de Kinetoplastida , Pez Cebra , Animales , Sistemas CRISPR-Cas/genética , ARN/genética , ARN Guía de Kinetoplastida/genética , ARN Mensajero/genética , Pez Cebra/genética
13.
BMC Plant Biol ; 10: 240, 2010 Nov 08.
Artículo en Inglés | MEDLINE | ID: mdl-21059227

RESUMEN

BACKGROUND: The economic importance of Solanaceae plant species is well documented and tomato has become a model for functional genomics studies. In plants, important processes are regulated by microRNAs (miRNA). DESCRIPTION: We describe here a data base integrating genetic map positions of miRNA-targeted genes, their expression profiles and their relations with quantitative fruit metabolic loci and yield associated traits. miSolRNA provides a metadata source to facilitate the construction of hypothesis aimed at defining physiological modes of action of regulatory process underlying the metabolism of the tomato fruit. CONCLUSIONS: The MiSolRNA database allows the simple extraction of metadata for the proposal of new hypothesis concerning possible roles of miRNAs in the regulation of tomato fruit metabolism. It permits i) to map miRNAs and their predicted target sites both on expressed (SGN-UNIGENES) and newly annotated sequences (BAC sequences released), ii) to co-locate any predicted miRNA-target interaction with metabolic QTL found in tomato fruits, iii) to retrieve expression data of target genes in tomato fruit along their developmental period and iv) to design further experiments for unresolved questions in complex trait biology based on the use of genetic materials that have been proven to be a useful tools for map-based cloning experiments in Solanaceae plant species.


Asunto(s)
Bases de Datos Genéticas , Frutas/genética , MicroARNs/genética , Solanum lycopersicum/genética , Secuencia de Bases , Sitios de Unión/genética , Biología Computacional/métodos , Frutas/crecimiento & desarrollo , Frutas/metabolismo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Almacenamiento y Recuperación de la Información , Solanum lycopersicum/crecimiento & desarrollo , Solanum lycopersicum/metabolismo , MicroARNs/metabolismo
14.
Dev Cell ; 54(6): 805-817.e7, 2020 09 28.
Artículo en Inglés | MEDLINE | ID: mdl-32768421

RESUMEN

Early embryonic development is driven exclusively by maternal gene products deposited into the oocyte. Although critical in establishing early developmental programs, maternal gene functions have remained elusive due to a paucity of techniques for their systematic disruption and assessment. CRISPR-Cas13 systems have recently been employed to degrade RNA in yeast, plants, and mammalian cell lines. However, no systematic study of the potential of Cas13 has been carried out in an animal system. Here, we show that CRISPR-RfxCas13d (CasRx) is an effective and precise system to deplete specific mRNA transcripts in zebrafish embryos. We demonstrate that zygotically expressed and maternally provided transcripts are efficiently targeted, resulting in a 76% average decrease in transcript levels and recapitulation of well-known embryonic phenotypes. Moreover, we show that this system can be used in medaka, killifish, and mouse embryos. Altogether, our results demonstrate that CRISPR-RfxCas13d is an efficient knockdown platform to interrogate gene function in animal embryos.


Asunto(s)
Sistemas CRISPR-Cas/genética , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas/genética , Edición Génica , Regulación del Desarrollo de la Expresión Génica/genética , Animales , Edición Génica/métodos , Células HEK293 , Humanos , Interferencia de ARN/fisiología , ARN Mensajero/genética
15.
BMC Plant Biol ; 9: 152, 2009 Dec 30.
Artículo en Inglés | MEDLINE | ID: mdl-20042107

RESUMEN

BACKGROUND: Micro RNAs (miRs) constitute a large group of endogenous small RNAs that have crucial roles in many important plant functions. Virus infection and transgenic expression of viral proteins alter accumulation and activity of miRs and so far, most of the published evidence involves post-transcriptional regulations. RESULTS: Using transgenic plants expressing a reporter gene under the promoter region of a characterized miR (P-miR164a), we monitored the reporter gene expression in different tissues and during Arabidopsis development. Strong expression was detected in both vascular tissues and hydathodes. P-miR164a activity was developmentally regulated in plants with a maximum expression at stages 1.12 to 5.1 (according to Boyes, 2001) along the transition from vegetative to reproductive growth. Upon quantification of P-miR164a-derived GUS activity after Tobacco mosaic virus Cg or Oilseed rape mosaic virus (ORMV) infection and after hormone treatments, we demonstrated that ORMV and gibberellic acid elevated P-miR164a activity. Accordingly, total mature miR164, precursor of miR164a and CUC1 mRNA (a miR164 target) levels increased after virus infection and interestingly the most severe virus (ORMV) produced the strongest promoter induction. CONCLUSION: This work shows for the first time that the alteration of miR pathways produced by viral infections possesses a transcriptional component. In addition, the degree of miR alteration correlates with virus severity since a more severe virus produces a stronger P-miR164a induction.


Asunto(s)
Arabidopsis/genética , Arabidopsis/virología , MicroARNs/metabolismo , Virus del Mosaico/fisiología , Regiones Promotoras Genéticas , Arabidopsis/metabolismo , Clonación Molecular , Biología Computacional , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes Reporteros , MicroARNs/genética , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/virología , Reguladores del Crecimiento de las Plantas/farmacología , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Plantas Modificadas Genéticamente/virología , ARN de Planta/genética
16.
Dev Cell ; 49(6): 867-881.e8, 2019 06 17.
Artículo en Inglés | MEDLINE | ID: mdl-31211993

RESUMEN

The awakening of the genome after fertilization is a cornerstone of animal development. However, the mechanisms that activate the silent genome after fertilization are poorly understood. Here, we show that transcriptional competency is regulated by Brd4- and P300-dependent histone acetylation in zebrafish. Live imaging of transcription revealed that genome activation, beginning at the miR-430 locus, is gradual and stochastic. We show that genome activation does not require slowdown of the cell cycle and is regulated through the translation of maternally inherited mRNAs. Among these, the enhancer regulators P300 and Brd4 can prematurely activate transcription and restore transcriptional competency when maternal mRNA translation is blocked, whereas inhibition of histone acetylation blocks genome activation. We conclude that P300 and Brd4 are sufficient to trigger genome-wide transcriptional competency by regulating histone acetylation on the first zygotic genes in zebrafish. This mechanism is critical for initiating zygotic development and developmental reprogramming.


Asunto(s)
Embrión no Mamífero/metabolismo , Desarrollo Embrionario , Regulación del Desarrollo de la Expresión Génica , Genoma , Proteínas de Pez Cebra/metabolismo , Pez Cebra/embriología , Cigoto/metabolismo , Animales , Proteína p300 Asociada a E1A/genética , Proteína p300 Asociada a E1A/metabolismo , Embrión no Mamífero/citología , Secuencias Reguladoras de Ácidos Nucleicos , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Activación Transcripcional , Transcriptoma , Proteínas de Pez Cebra/genética , Cigoto/citología
18.
Cold Spring Harb Protoc ; 2016(10)2016 10 03.
Artículo en Inglés | MEDLINE | ID: mdl-27698232

RESUMEN

This protocol describes how to generate and genotype mutants using an optimized CRISPR-Cas9 genome-editing system in zebrafish (CRISPRscan). Because single guide RNAs (sgRNAs) have variable efficiency when targeting specific loci, our protocol starts by explaining how to use the web tool CRISPRscan to design highly efficient sgRNAs. The CRISPRscan algorithm is based on the results of an integrated analysis of more than 1000 sgRNAs in zebrafish, which uncovered highly predictive factors that influence Cas9 activity. Next, we describe how to easily generate sgRNAs in vitro, which can then be injected in vivo to target specific loci. The use of highly efficient sgRNAs can lead to biallelic mutations in the injected embryos, causing lethality. We explain how targeting Cas9 to the germline increases viability by reducing somatic mutations. Finally, we combine two methods to identify F1 heterozygous fish carrying the desired mutations: (i) Mut-Seq, a method based on high-throughput sequencing to detect F0 founder fish; and (ii) a polymerase chain reaction-based fragment analysis method that identifies F1 heterozygous fish characterized by Mut-Seq. In summary, this protocol includes the steps to generate and characterize mutant zebrafish lines using the CRISPR-Cas9 genome engineering system.


Asunto(s)
Sistemas CRISPR-Cas , Edición Génica/métodos , Pez Cebra/genética , Animales , Biología Computacional/métodos , Endonucleasas/metabolismo , Técnicas de Genotipaje/métodos , ARN Guía de Kinetoplastida/metabolismo
19.
Cold Spring Harb Protoc ; 2016(10)2016 10 03.
Artículo en Inglés | MEDLINE | ID: mdl-27698246

RESUMEN

The CRISPR-Cas9 system uncovered in bacteria has emerged as a powerful genome-editing technology in eukaryotic cells. It consists of two components-a single guide RNA (sgRNA) that directs the Cas9 endonuclease to a complementary DNA target site. Efficient targeting of individual genes requires highly active sgRNAs. Recent efforts have made significant progress in understanding the sequence features that increase sgRNA activity. In this introduction, we highlight advancements in the field of CRISPR-Cas9 targeting and discuss our web tool CRISPRscan, which predicts the targeting activity of sgRNAs and improves the efficiency of the CRISPR-Cas9 system for in vivo genome engineering.


Asunto(s)
Sistemas CRISPR-Cas , Edición Génica/métodos , Endonucleasas/metabolismo , ARN Guía de Kinetoplastida/metabolismo , Recombinación Genética
20.
Mol Plant Pathol ; 17(3): 317-29, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26033697

RESUMEN

Sweet orange (Citrus sinensis), one of the most important fruit crops worldwide, may suffer from disease symptoms induced by virus infections, thus resulting in dramatic economic losses. Here, we show that the infection of sweet orange plants with two isolates of Citrus psorosis virus (CPsV) expressing different symptomatology alters the accumulation of a set of endogenous microRNAs (miRNAs). Within these miRNAs, miR156, miR167 and miR171 were the most down-regulated, with almost a three-fold reduction in infected samples. This down-regulation led to a concomitant up-regulation of some of their targets, such as Squamosa promoter-binding protein-like 9 and 13, as well as Scarecrow-like 6. The processing of miRNA precursors, pre-miR156 and pre-miR171, in sweet orange seems to be affected by the virus. For instance, virus infection increases the level of unprocessed precursors, which is accompanied by a concomitant decrease in mature species accumulation. miR156a primary transcript accumulation remained unaltered, thus strongly suggesting a processing deregulation for this transcript. The co-immunoprecipitation of viral 24K protein with pre-miR156a or pre-miR171a suggests that the alteration in the processing of these precursors might be caused by a direct or indirect interaction with this particular viral protein. This result is also consistent with the nuclear localization of both miRNA precursors and the CPsV 24K protein. This study contributes to the understanding of the manner in which a virus can alter host regulatory mechanisms, particularly miRNA biogenesis and target expression.


Asunto(s)
Citrus/virología , Regulación Viral de la Expresión Génica , MicroARNs/genética , Virus de Plantas/genética , Precursores del ARN/genética , Procesamiento Postranscripcional del ARN/genética , Proteínas Virales/metabolismo , Secuencia Conservada/genética , Proteínas Fluorescentes Verdes/metabolismo , Inmunoprecipitación , MicroARNs/metabolismo , Enfermedades de las Plantas/virología , Plantas Modificadas Genéticamente , Unión Proteica , Precursores del ARN/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Nicotiana/genética
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