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2.
Biomolecules ; 13(2)2023 01 19.
Article in English | MEDLINE | ID: mdl-36830576

ABSTRACT

Primary transcripts of microRNAs (pri-miRNAs) were initially defined as long non-coding RNAs that host miRNAs further processed by the microRNA processor complex. A few years ago, however, it was discovered in plants that pri-miRNAs actually contain functional open reading frames (sORFs) that translate into small peptides called miPEPs, for microRNA-encoded peptides. Initially detected in Arabidopsis thaliana and Medicago truncatula, recent studies have revealed the presence of miPEPs in other pri-miRNAs as well as in other species ranging from various plant species to animals. This suggests that miPEP numbers remain largely underestimated and that they could be a common signature of pri-miRNAs. Here we present the most recent advances in miPEPs research and discuss how their discovery has broadened our vision of the regulation of gene expression by miRNAs, and how miPEPs could be interesting tools in sustainable agriculture or the treatment of certain human diseases.


Subject(s)
Arabidopsis , MicroRNAs , RNA, Long Noncoding , Humans , Animals , MicroRNAs/genetics , Peptides/genetics , Plants/genetics , Arabidopsis/genetics , Gene Expression Regulation, Plant
3.
Nat Commun ; 14(1): 254, 2023 01 17.
Article in English | MEDLINE | ID: mdl-36650156

ABSTRACT

The current agriculture main challenge is to maintain food production while facing multiple threats such as increasing world population, temperature increase, lack of agrochemicals due to health issues and uprising of weeds resistant to herbicides. Developing novel, alternative, and safe methods is hence of paramount importance. Here, we show that complementary peptides (cPEPs) from any gene can be designed to target specifically plant coding genes. External application of synthetic peptides increases the abundance of the targeted protein, leading to related phenotypes. Moreover, we provide evidence that cPEPs can be powerful tools in agronomy to improve plant traits, such as growth, resistance to pathogen or heat stress, without the needs of genetic approaches. Finally, by combining their activity they can also be used to reduce weed growth.


Subject(s)
Agrochemicals , Weed Control , Agrochemicals/pharmacology , Herbicide Resistance/genetics , Plant Weeds/genetics , Peptides , Crops, Agricultural/genetics
4.
Front Cell Dev Biol ; 10: 901351, 2022.
Article in English | MEDLINE | ID: mdl-35721519

ABSTRACT

Recent studies have shown that hundreds of small proteins were occulted when protein-coding genes were annotated. These proteins, called alternative proteins, have failed to be annotated notably due to the short length of their open reading frame (less than 100 codons) or the enforced rule establishing that messenger RNAs (mRNAs) are monocistronic. Several alternative proteins were shown to be biologically active molecules and seem to be involved in a wide range of biological functions. However, genome-wide exploration of the alternative proteome is still limited to a few species. In the present article, we describe a deep peptidomics workflow which enabled the identification of 401 alternative proteins in Drosophila melanogaster. Subcellular localization, protein domains, and short linear motifs were predicted for 235 of the alternative proteins identified and point toward specific functions of these small proteins. Several alternative proteins had approximated abundances higher than their canonical counterparts, suggesting that these alternative proteins are actually the main products of their corresponding genes. Finally, we observed 14 alternative proteins with developmentally regulated expression patterns and 10 induced upon the heat-shock treatment of embryos, demonstrating stage or stress-specific production of alternative proteins.

5.
Int J Mol Sci ; 23(10)2022 May 20.
Article in English | MEDLINE | ID: mdl-35628573

ABSTRACT

MicroRNAs (miRNAs) are small regulatory non-coding RNAs, resulting from the cleavage of long primary transcripts (pri-miRNAs) in the nucleus by the Microprocessor complex generating precursors (pre-miRNAs) that are then exported to the cytoplasm and processed into mature miRNAs. Some miRNAs are hosted in pri-miRNAs annotated as long non-coding RNAs (lncRNAs) and defined as MIRHGs (for miRNA Host Genes). However, several lnc pri-miRNAs contain translatable small open reading frames (smORFs). If smORFs present within lncRNAs can encode functional small peptides, they can also constitute cis-regulatory elements involved in lncRNA decay. Here, we investigated the possible involvement of smORFs in the regulation of lnc pri-miRNAs in Human and Drosophila, focusing on pri-miRNAs previously shown to contain translatable smORFs. We show that smORFs regulate the expression levels of human pri-miR-155 and pri-miR-497, and Drosophila pri-miR-8 and pri-miR-14, and also affect the expression and activity of their associated miRNAs. This smORF-dependent regulation is independent of the nucleotidic and amino acidic sequences of the smORFs and is sensitive to the ribosome-stalling drug cycloheximide, suggesting the involvement of translational events. This study identifies smORFs as new cis-acting elements involved in the regulation of pri-miRNAs and miRNAs expression, in both Human and Drosophila melanogaster.


Subject(s)
MicroRNAs , RNA, Long Noncoding , RNA, Small Untranslated , Animals , Drosophila/genetics , Drosophila/metabolism , Drosophila melanogaster/genetics , Drosophila melanogaster/metabolism , Humans , MicroRNAs/genetics , MicroRNAs/metabolism , Open Reading Frames/genetics
6.
Cell Rep ; 38(6): 110339, 2022 02 08.
Article in English | MEDLINE | ID: mdl-35139385

ABSTRACT

MicroRNAs (miRNAs) are transcribed as long primary transcripts (pri-miRNAs) by RNA polymerase II. Plant pri-miRNAs encode regulatory peptides called miPEPs, which specifically enhance the transcription of the pri-miRNA from which they originate. However, paradoxically, whereas miPEPs have been identified in different plant species, they are poorly conserved, raising the question of the mechanisms underlying their specificity. To address this point, we identify and re-annotate multiple Arabidopsis thaliana pri-miRNAs in order to identify ORF encoding miPEPs. The study of several identified miPEPs in different species show that non-conserved miPEPs are only active in their plant of origin, whereas conserved ones are active in different species. Finally, we find that miPEP activity relies on the presence of its own miORF, explaining both the lack of selection pressure on miPEP sequence and the ability for non-conserved peptides to play a similar role, i.e., to activate the expression of their corresponding miRNA.


Subject(s)
Arabidopsis/metabolism , Gene Expression Regulation, Plant/genetics , MicroRNAs/metabolism , Peptides/metabolism , Open Reading Frames/genetics , Plants/genetics
7.
J Hum Evol ; 162: 103092, 2022 01.
Article in English | MEDLINE | ID: mdl-34839228

ABSTRACT

Previous studies have suggested that the Lower-to-Middle Paleolithic transition was associated with the earliest Neanderthals, but recent research has established that the oldest Neanderthal fossils and the first signs of their technologies and behavior appear from MIS 11 or possibly earlier. To understand these changes, re-evaluation of the evidence is necessary to determine if this transition corresponds to a progressive evolution rather than abrupt change. Orgnac 3 is a key and appropriate site to study this research context. Located in southern France, it yields a long stratigraphic sequence testifying the evolution of technical and subsistence behaviors of pre-Neanderthal human groups during a Middle Pleistocene interglacial-glacial cycle. In this article, a new chronological framework is provided for the sequence based on results of dating methods applied to various types of geological materials. Speleothems and volcanic minerals, dated in previous studies by U-series and 40Ar/39Ar, respectively, show periods of calcitic crystallization and regional volcanic activity. Other materials, such as heated flints and herbivore teeth, are directly related to evidence of anthropogenic activities and are analyzed in the present work by trapped-charge dating methods such as thermoluminescence and electron spin resonance combined with uranium series (ESR/U-series). The new thermoluminescence and ESR/U-series dates confirm the attribution of the Orgnac 3 stratigraphic sequence to the MIS 10-MIS 8 period and are discussed in relation to paleoenvironmental data derived from bioarchaeological studies. The paleoanthropological levels, including the emergence of Levallois technology, are dated to ca. 275 ka (early MIS 8) and appear coeval to a wet and temperate period recorded locally, the Amargiers interstadial, defined in the regional palynological records. The implications of this reassessed chronology for the archaeological assemblages are discussed in the wider context of behavioral innovations from MIS 11 onward and their establishment in subsequent periods.


Subject(s)
Neanderthals , Tooth , Animals , Archaeology , Fossils , France , Humans , Technology
9.
Int J Mol Sci ; 22(7)2021 Mar 26.
Article in English | MEDLINE | ID: mdl-33810468

ABSTRACT

Some miRNAs are located in RNA precursors (pri-miRNAs) annotated as long non-coding (lncRNAs) due to absence of long open reading frames (ORFs). However, recent studies have shown that some lnc pri-miRNAs encode peptides called miPEPs (miRNA-encoded peptides). Initially discovered in plants, three miPEPs have also been identified in humans. Herein, we found that a dozen human pri-miRNAs potentially encode miPEPs, as revealed by ribosome profiling and proteomic databases survey. So far, the only known function of plant miPEPs is to enhance the transcription of their own pri-miRNAs, thereby increasing the level and activity of their associated miRNAs and downregulating the expression of their target genes. To date, in humans, only miPEP133 was shown to promote a positive autoregulatory loop. We investigated whether other human miPEPs are also involved in regulating the expression of their miRNAs by studying miPEP155, encoded by the lnc MIR155HG, miPEP497, a sORF-encoded peptide within lnc MIR497HG, and miPEP200a, encoded by the pri-miRNA of miR-200a/miR-200b. We show that overexpression of these miPEPs is unable to impact the expression/activity of their own pri-miRNA/miRNAs in humans, indicating that the positive feedback regulation observed with plant miPEPs and human miPEP133 is not a general rule of human miPEP function.


Subject(s)
Gene Expression Regulation, Neoplastic , Gene Expression Regulation, Plant , Gene Expression Regulation , MicroRNAs/genetics , HeLa Cells , Humans , MicroRNAs/chemistry , Open Reading Frames , PC-3 Cells , Peptides/chemistry , Proteomics , RNA Precursors/genetics , RNA Processing, Post-Transcriptional
10.
Genome Biol ; 22(1): 118, 2021 04 23.
Article in English | MEDLINE | ID: mdl-33892772

ABSTRACT

BACKGROUND: Recent genome-wide studies of many species reveal the existence of a myriad of RNAs differing in size, coding potential and function. Among these are the long non-coding RNAs, some of them producing functional small peptides via the translation of short ORFs. It now appears that any kind of RNA presumably has a potential to encode small peptides. Accordingly, our team recently discovered that plant primary transcripts of microRNAs (pri-miRs) produce small regulatory peptides (miPEPs) involved in auto-regulatory feedback loops enhancing their cognate microRNA expression which in turn controls plant development. Here we investigate whether this regulatory feedback loop is present in Drosophila melanogaster. RESULTS: We perform a survey of ribosome profiling data and reveal that many pri-miRNAs exhibit ribosome translation marks. Focusing on miR-8, we show that pri-miR-8 can produce a miPEP-8. Functional assays performed in Drosophila reveal that miPEP-8 affects development when overexpressed or knocked down. Combining genetic and molecular approaches as well as genome-wide transcriptomic analyses, we show that miR-8 expression is independent of miPEP-8 activity and that miPEP-8 acts in parallel to miR-8 to regulate the expression of hundreds of genes. CONCLUSION: Taken together, these results reveal that several Drosophila pri-miRs exhibit translation potential. Contrasting with the mechanism described in plants, these data shed light on the function of yet undescribed primary-microRNA-encoded peptides in Drosophila and their regulatory potential on genome expression.


Subject(s)
Drosophila/genetics , Gene Expression Regulation , MicroRNAs/genetics , Peptides/genetics , Animals , Female , Gene Expression Profiling , Male , MicroRNAs/chemistry , Mutation , Nucleic Acid Conformation , Open Reading Frames , Phenotype , Protein Biosynthesis , RNA Interference , RNA, Long Noncoding
11.
Curr Opin Chem Biol ; 60: 122-130, 2021 02.
Article in English | MEDLINE | ID: mdl-33401134

ABSTRACT

Short open reading frame (sORF)-encoded polypeptides (SEPs) have recently emerged as key regulators of major cellular processes. Computational methods for the annotation of sORFs combined with transcriptomics and ribosome profiling approaches predicted the existence of tens of thousands of SEPs across the kingdom of life. Although, we still lack unambiguous evidence for most of them. The method of choice to validate the expression of SEPs is mass spectrometry (MS)-based peptidomics. Peptides are less abundant than proteins, which tends to hinder their detection. Therefore, optimization and enrichment methods are necessary to validate the existence of SEPs. In this article, we discuss the challenges for the detection of SEPs by MS and recent developments of biochemical approaches applied to the study of these peptides. We detail the advances made in the different key steps of a typical peptidomics workflow and highlight possible alternatives that have not been explored yet.


Subject(s)
Mass Spectrometry , Open Reading Frames/genetics , Peptides/genetics , Peptides/metabolism , Proteomics/methods , Workflow
12.
Trends Plant Sci ; 26(2): 132-141, 2021 02.
Article in English | MEDLINE | ID: mdl-33036916

ABSTRACT

The importance of microorganisms in plant development, nutrition, and stress resistance is unquestioned and has led to a more holistic approach of plant-microbe interactions, under the holobiont concept. The structure of the plant microbiota is often described as host driven, especially in the rhizosphere, where microbial communities are shaped by diverse rhizodeposits. Gradually, this anthropogenic vision is fading and being replaced by the idea that plants and microorganisms co-shape the plant microbiota. Through coevolution, plants and microbes have developed cross-kingdom communication channels. Here, we propose that miRNAs are crucial mediators of plant-microbe interactions and microbiota shaping in the rhizosphere. Moreover, we suggest, as an alternative to generally unsuccessful strategies based on microbial inoculants, miRNAs as a promising tool for novel holobiont engineering.


Subject(s)
MicroRNAs , Microbiota , MicroRNAs/genetics , Plant Development , Plants/genetics , Rhizosphere , Soil Microbiology
13.
Int J Mol Sci ; 21(7)2020 Mar 25.
Article in English | MEDLINE | ID: mdl-32218176

ABSTRACT

MiPEPs are short natural peptides encoded by microRNAs in plants. Exogenous application of miPEPs increases the expression of their corresponding miRNA and, consequently, induces consistent phenotypical changes. Therefore, miPEPs carry huge potential in agronomy as gene regulators that do not require genome manipulation. However, to this end, it is necessary to know their mode of action, including where they act and how they enter the plants. Here, after analyzing the effect of Arabidopsis thaliana miPEP165a on root and aerial part development, we followed the internalization of fluorescent-labelled miPEP165a into roots and compared its uptake into endocytosis-altered mutants to that observed in wild-type plants treated or not with endocytosis inhibitors. The results show that entry of miPEP165a involves both a passive diffusion at the root apex and endocytosis-associated internalization in the differentiation and mature zones. Moreover, miPEP165a is unable to enter the central cylinder and does not migrate from the roots to the aerial part of the plant, suggesting that miPEPs have no systemic effect.


Subject(s)
Arabidopsis/drug effects , Endocytosis , Arabidopsis/cytology , Arabidopsis/metabolism , Biological Transport , Cell Division/drug effects , Diffusion , Endocytosis/drug effects , Phenotype , Plant Roots/cytology , Plant Roots/drug effects , Plant Roots/metabolism , Plants, Genetically Modified
14.
Cell Host Microbe ; 21(1): 106-112, 2017 Jan 11.
Article in English | MEDLINE | ID: mdl-28041928

ABSTRACT

Arbuscular mycorrhizal (AM) symbiosis associates most plants with fungi of the phylum Glomeromycota. The fungus penetrates into roots and forms within cortical cell branched structures called arbuscules for nutrient exchange. We discovered that miR171b has a mismatched cleavage site and is unable to downregulate the miR171 family target gene, LOM1 (LOST MERISTEMS 1). This mismatched cleavage site is conserved among plants that establish AM symbiosis, but not in non-mycotrophic plants. Unlike other members of the miR171 family, miR171b stimulates AM symbiosis and is expressed specifically in root cells that contain arbuscules. MiR171b protects LOM1 from negative regulation by other miR171 family members. These findings uncover a unique mechanism of positive post-transcriptional regulation of gene expression by miRNAs and demonstrate its relevance for the establishment of AM symbiosis.


Subject(s)
Glomeromycota/growth & development , Medicago truncatula/genetics , Medicago truncatula/microbiology , MicroRNAs/genetics , Mycorrhizae/growth & development , Nicotiana/genetics , Nicotiana/microbiology , Base Sequence , Gene Expression Regulation, Plant , Plant Roots/microbiology , Symbiosis/physiology
15.
Am J Phys Anthropol ; 162(1): 186-190, 2017 01.
Article in English | MEDLINE | ID: mdl-27652578

ABSTRACT

OBJECTIVES: Handedness is an important marker for lateralization of humans in the modern and fossil record. For the most part, Neandertals and their ancestors are strongly right-handed. We describe a single tooth from a Neandertal level at Vergisson 4 (Vg 4-83). This left upper central incisor shows all the features typical of Neandertal incisors. It also exhibits a predominance of left-handed striations. METHODS: Striations on the incisor's labial surface were mapped at 20x magnification using Photoshop. Angulations of the striations were determined from their deviation from the maximum mesio-distal line and were analyzed using NIH's freeware, Image J. RESULTS: Of the 60 labial surface striations, Vg 4-83 shows a strong predominance of left-handed striations (46; 76.7%), which are statistically significantly different (p < .001 with a two-tailed chi2 test) from the small number (3) of right-handed striations. DISCUSSION: The identification of another left-handed Neandertal adds to our understanding about handedness variation in this fossil hominin. Given the high frequency of right-handed Neandertals, the 90: 10 modern ratio is still preserved in this group.


Subject(s)
Functional Laterality/physiology , Incisor/pathology , Neanderthals/anatomy & histology , Animals , Anthropology, Physical , Fossils , France
16.
New Phytol ; 213(3): 1124-1132, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27748948

ABSTRACT

Root colonization by arbuscular mycorrhizal (AM) fungi is a complex and finely tuned process. Previous studies have shown that, among other plant hormones, auxin plays a role in this process but the specific involvement of Aux/IAAs, the key regulators of auxin responses, is still unknown. In this study, we addressed the role of the tomato Sl-IAA27 during AM symbiosis by using Sl-IAA27-RNAi and pSL-IAA27::GUS stable tomato lines. The data show that Sl-IAA27 expression is up-regulated by the AM fungus and that silencing of Sl-IAA27 has a negative impact on AM colonization. Sl-IAA27-silencing resulted in down-regulation of three genes involved in strigolactone synthesis, NSP1, D27 and MAX1, and treatment of Sl-IAA27-silenced plants with the strigolactone analog GR24 complemented their mycorrhizal defect phenotype. Overall, the study identified an Aux/IAA gene as a new component of the signaling pathway controlling AM fungal colonization in tomato. This gene is proposed to control strigolactone biosynthesis via the regulation of NSP1.


Subject(s)
Glomeromycota/physiology , Heterocyclic Compounds, 3-Ring/metabolism , Lactones/metabolism , Mycorrhizae/physiology , Plant Proteins/metabolism , Solanum lycopersicum/metabolism , Solanum lycopersicum/microbiology , Gene Expression Regulation, Plant , Genetic Complementation Test , Phenotype , Plant Roots/metabolism , RNA Interference
17.
Front Plant Sci ; 7: 1704, 2016.
Article in English | MEDLINE | ID: mdl-27899928

ABSTRACT

Arbuscular mycorrhizal (AM) symbiosis is an intimate and ancient symbiosis found between most of terrestrial plants and fungi from the Glomeromycota family. Later during evolution, the establishment of the nodulation between legume plants and soil bacteria known as rhizobia, involved several genes of the signaling pathway previously implicated for AM symbiosis. For the past years, the identification of the genes belonging to this Common Symbiotic Signaling Pathway have been mostly done on nodulation. Among the different genes already well identified as required for nodulation, we focused our attention on the involvement of Nodule Inception (NIN) in AM symbiosis. We show here that NIN expression is induced during AM symbiosis, and that the Medicago truncatula nin mutant is less colonized than the wild-type M. truncatula strain. Moreover, nin mutant displays a defect in the ability to be infected by the fungus Rhizophagus irregularis. This work brings a new evidence of the common genes involved in overlapping signaling pathways of both nodulation and in AM symbiosis.

18.
New Phytol ; 212(1): 22-35, 2016 10.
Article in English | MEDLINE | ID: mdl-27292927

ABSTRACT

Contents 22 I. 22 II. 24 III. 25 IV. 27 V. 29 VI. 10 31 References 32 SUMMARY: Plants have evolved a remarkable faculty of adaptation to deal with various and changing environmental conditions. In this context, the roots have taken over nutritional aspects and the root system architecture can be modulated in response to nutrient availability or biotic interactions with soil microorganisms. This adaptability requires a fine tuning of gene expression. Indeed, root specification and development are highly complex processes requiring gene regulatory networks involved in hormonal regulations and cell identity. Among the different molecular partners governing root development, microRNAs (miRNAs) are key players for the fast regulation of gene expression. miRNAs are small RNAs involved in most developmental processes and are required for the normal growth of organisms, by the negative regulation of key genes, such as transcription factors and hormone receptors. Here, we review the known roles of miRNAs in root specification and development, from the embryonic roots to the establishment of root symbioses, highlighting the major roles of miRNAs in these processes.


Subject(s)
MicroRNAs/metabolism , Plant Roots/anatomy & histology , Plant Roots/genetics , Meristem/genetics , MicroRNAs/genetics , Plant Root Nodulation/genetics , Plant Roots/embryology , Symbiosis/genetics
20.
RNA Biol ; 12(11): 1178-80, 2015.
Article in English | MEDLINE | ID: mdl-26400469

ABSTRACT

MicroRNAs (miRNAs) are short RNA molecules negatively regulating the expression of many important genes in plants and animals. We have recently shown that plant primary transcripts of miRNAs encode peptides (miPEPs) able to increase specifically the transcription of their associated miRNA.(1) We discuss here the possibility of using miPEPs as a new tool for functional analysis of single members of miRNA families in plants, including in non-model plants, that could avoid transgenic transformation and minimize artifactual interpretation. We also raise several fundamental and crucial questions that need to be address for a deeper understanding of the cellular and molecular mechanisms underlining the regulatory activity of miPEPs.


Subject(s)
MicroRNAs/genetics , Peptides/genetics , Plants/genetics , Gene Expression Regulation, Plant , Open Reading Frames , Peptides/metabolism , Plants/metabolism
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