Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 14 de 14
Filter
Add more filters










Publication year range
1.
iScience ; 26(10): 107723, 2023 Oct 20.
Article in English | MEDLINE | ID: mdl-37692287

ABSTRACT

Splicing of transcripts is catalyzed by the spliceosome, a mega-complex consisting of hundreds of proteins and five snRNAs, which employs direct interactions. When U1 snRNA forms high-affinity binding, namely more than eight base pairs, with the 5'SS, the result is usually a suppressing effect on the splicing activity. This likely occurs due to the inefficient unwinding of U1/5'SS base-pairing or other regulatory obstructions. Here, we show in vitro and in patient-derived cell lines that pre-microRNAs can modulate the splicing reaction by interacting with U1 snRNA. This leads to reduced binding affinity to the 5'SS, and hence promotes the inclusion of exons containing 5'SS, despite sequence-based high affinity to U1. Application of the mechanism resulted in correction of the splicing defect in the disease-causing VCAN gene from an individual with Wagner syndrome. This pre-miRNA/U1 interaction can regulate the expression of alternatively spliced exons, thus extending the scope of mechanisms regulating splicing.

2.
Cells ; 12(2)2023 01 06.
Article in English | MEDLINE | ID: mdl-36672174

ABSTRACT

Rapid responses to changes within subcellular compartments of highly polarized cells, such as neuron axons, depend on local translation and post-transcriptional regulation. The mechanism by which microRNAs (miRNAs) regulate this process is not fully understood. Here, using live cell imaging and RNA sequencing analysis, we demonstrated how miRNAs can differentially control hundreds of transcripts at the subcellular level. We demonstrated that the seed match length of the miRNA target-sequence regulates both mRNA stability and protein translation rates. While longer seed matches have an increased inhibitory effect, transcriptome analysis did not reveal differences in seed match length between axonal and somata mRNAs of motor neurons. However, mRNA variants with longer 3'UTR are enriched in axons and contain multiple repeats of specific miRNA target sequences. Finally, we demonstrated that the long 3'UTR mRNA variant of the motor protein Kif5b is enriched explicitly in motor neuron axons and contains multiple sequence repeats for binding miR-129-5p. This subsequently results in the differential post-transcriptional regulation of kif5b and its synthesis in axons. Thus, we suggest that the number of miRNA binding sites at the 3'UTR of the mRNA, rather than the miRNA seed match length, regulates the axonal transcriptome.


Subject(s)
MicroRNAs , 3' Untranslated Regions/genetics , MicroRNAs/genetics , MicroRNAs/metabolism , Axons/metabolism , Binding Sites , Carrier Proteins/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism
3.
Nat Commun ; 12(1): 6914, 2021 11 25.
Article in English | MEDLINE | ID: mdl-34824257

ABSTRACT

Mislocalization of the predominantly nuclear RNA/DNA binding protein, TDP-43, occurs in motor neurons of ~95% of amyotrophic lateral sclerosis (ALS) patients, but the contribution of axonal TDP-43 to this neurodegenerative disease is unclear. Here, we show TDP-43 accumulation in intra-muscular nerves from ALS patients and in axons of human iPSC-derived motor neurons of ALS patient, as well as in motor neurons and neuromuscular junctions (NMJs) of a TDP-43 mislocalization mouse model. In axons, TDP-43 is hyper-phosphorylated and promotes G3BP1-positive ribonucleoprotein (RNP) condensate assembly, consequently inhibiting local protein synthesis in distal axons and NMJs. Specifically, the axonal and synaptic levels of nuclear-encoded mitochondrial proteins are reduced. Clearance of axonal TDP-43 or dissociation of G3BP1 condensates restored local translation and resolved TDP-43-derived toxicity in both axons and NMJs. These findings support an axonal gain of function of TDP-43 in ALS, which can be targeted for therapeutic development.


Subject(s)
Axons/metabolism , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/metabolism , Inhibition, Psychological , Mitochondrial Proteins/metabolism , Neuromuscular Junction/metabolism , Amyotrophic Lateral Sclerosis/drug therapy , Animals , C9orf72 Protein/genetics , DNA Helicases , DNA-Binding Proteins/genetics , DNA-Binding Proteins/pharmacology , Disease Models, Animal , Female , Humans , Induced Pluripotent Stem Cells , Mice , Mice, Inbred C57BL , Mitochondria/drug effects , Mitochondrial Proteins/chemistry , Mitochondrial Proteins/genetics , Motor Neurons , Neurodegenerative Diseases/drug therapy , Neuromuscular Junction/drug effects , Neurons/drug effects , Neurons/metabolism , Neurons, Efferent , Phosphorylation , Poly-ADP-Ribose Binding Proteins , RNA Helicases , RNA Recognition Motif Proteins
4.
J Perinat Med ; 49(9): 1071-1083, 2021 Nov 25.
Article in English | MEDLINE | ID: mdl-34114389

ABSTRACT

OBJECTIVES: Preeclampsia is a dangerous pregnancy complication. The source of preeclampsia is unknown, though the placenta is believed to have a central role in its pathogenesis. An association between maternal infection and preeclampsia has been demonstrated, yet the involvement of the placental microbiome in the etiology of preeclampsia has not been determined. In this study, we examined whether preeclampsia is associated with an imbalanced microorganism composition in the placenta. METHODS: To this end, we developed a novel method for the identification of bacteria/viruses based on sequencing of small non-coding RNA, which increases the microorganism-to-host ratio, this being a major challenge in microbiome methods. We validated the method on various infected tissues and demonstrated its efficiency in detecting microorganisms in samples with extremely low bacterial/viral biomass. We then applied the method to placenta specimens from preeclamptic and healthy pregnancies. Since the placenta is a remarkably large and heterogeneous organ, we explored the bacterial and viral RNA at each of 15 distinct locations. RESULTS: Bacterial RNA was detected at all locations and was consistent with previous studies of the placental microbiome, though without significant differences between the preeclampsia and control groups. Nevertheless, the bacterial RNA composition differed significantly between various areas of the placenta. Viral RNA was detected in extremely low quantities, below the threshold of significance, thus viral abundance could not be determined. CONCLUSIONS: Our results suggest that the bacterial and viral abundance in the placenta may have only limited involvement in the pathogenesis of preeclampsia. The evidence of a heterogenic bacterial RNA composition in the various placental locations warrants further investigation to capture the true nature of the placental microbiome.


Subject(s)
Microbiota/genetics , Placenta/microbiology , Pre-Eclampsia , RNA, Bacterial , RNA, Viral , Sequence Analysis, RNA , Adult , Bacteria/classification , Bacteria/isolation & purification , Correlation of Data , Female , Humans , Outcome Assessment, Health Care , Placenta/pathology , Pre-Eclampsia/blood , Pre-Eclampsia/diagnosis , Pre-Eclampsia/microbiology , Pregnancy , RNA, Bacterial/analysis , RNA, Bacterial/isolation & purification , RNA, Untranslated/analysis , RNA, Untranslated/isolation & purification , RNA, Viral/analysis , RNA, Viral/isolation & purification , Reproducibility of Results , Sequence Analysis, RNA/methods , Sequence Analysis, RNA/statistics & numerical data , Specimen Handling/methods
5.
PLoS One ; 14(1): e0211111, 2019.
Article in English | MEDLINE | ID: mdl-30682089

ABSTRACT

HIV-1 is the causative agent of AIDS (Autoimmune Deficiency Syndrome). HIV-1 infection results in systemic CD4+ T cell depletion, thereby impairing cell-mediated immunity. MicroRNAs are short (~22 nucleotides long), endogenous single-stranded RNA molecules that regulate gene expression by binding to the 3' untranslated regions (3' UTR) of mRNA transcripts. The relation between HIV-1 infection and human miRNA expression profile has been previously investigated, and studies have shown that the virus can alter miRNA expression and vice versa. Here, we broaden the understanding of the HIV-1 infection process, and show that miRNA-186, 210 and 222 are up-regulated following HIV-1 infection of human Sup-T1 cells. As a result, the host miRNA target genes: Dicer1 (Double-Stranded RNA-Specific Endoribonuclease), HRB (HIV-1 Rev-binding protein) and HIV-EP2 (Human Immunodeficiency Virus Type I Enhancer Binding Protein 2), are down-regulated. Moreover, testing the miRNA-gene anti- correlation on the Jurkat and the HeLa-MAGI cell lines demonstrated the ability of the miRNAs to down-regulate viral expression as well. To conclude, we found that human miR-186, 210 and 222 directly regulate the human genes Dicer1, HRB and HIV-EP2, thus may be filling key roles during HIV-1 replication and miRNA biogenesis. This finding may contribute to the development of new therapeutic strategies.


Subject(s)
DEAD-box RNA Helicases/biosynthesis , DNA-Binding Proteins/biosynthesis , Down-Regulation , HIV Infections/metabolism , HIV-1/physiology , MicroRNAs/metabolism , Nuclear Pore Complex Proteins/biosynthesis , RNA-Binding Proteins/biosynthesis , Ribonuclease III/biosynthesis , Transcription Factors/biosynthesis , Virus Replication/physiology , HeLa Cells , Humans , Jurkat Cells , MCF-7 Cells
6.
Sci Rep ; 8(1): 3401, 2018 02 21.
Article in English | MEDLINE | ID: mdl-29467498

ABSTRACT

Preeclampsia is one of the most dangerous pregnancy complications, and the leading cause of maternal and perinatal mortality and morbidity. Although the clinical symptoms appear late, its origin is early, and hence detection is feasible already at the first trimester. In the current study, we investigated the abundance of circulating small non-coding RNAs in the plasma of pregnant women in their first trimester, seeking transcripts that best separate the preeclampsia samples from those of healthy pregnant women. To this end, we performed small non-coding RNAs sequencing of 75 preeclampsia and control samples, and identified 25 transcripts that were differentially expressed between preeclampsia and the control groups. Furthermore, we utilized those transcripts and created a pipeline for a supervised classification of preeclampsia. Our pipeline generates a logistic regression model using a 5-fold cross validation on numerous random partitions into training and blind test sets. Using this classification procedure, we achieved an average AUC value of 0.86. These findings suggest the predictive value of circulating small non-coding RNA in the first trimester, warranting further examination, and lay the foundation for producing a novel early non-invasive diagnostic tool for preeclampsia, which could reduce the life-threatening risk for both the mother and fetus.


Subject(s)
Pre-Eclampsia/blood , Pre-Eclampsia/diagnosis , RNA, Small Untranslated/blood , Adult , Biomarkers/blood , Case-Control Studies , Early Diagnosis , Female , Gestational Age , Humans , Infant, Newborn , Male , Pregnancy , Pregnancy Outcome , Pregnancy Trimester, First/blood , Pregnancy Trimester, Second/blood , Prospective Studies , Risk Factors
8.
Front Mol Neurosci ; 10: 355, 2017.
Article in English | MEDLINE | ID: mdl-29163031

ABSTRACT

Background: Selective serotonin reuptake inhibitor (SSRI) antidepressant drugs are the first-line of treatment for major depressive disorder (MDD) but are effective in <70% of patients. Our earlier genome-wide studies indicated that two genes encoding for cell adhesion proteins, close homolog of L1 (CHL1) and integrin beta-3 (ITGB3), and microRNAs, miR-151a-3p and miR-221/222, are implicated in the variable sensitivity and response of human lymphoblastoid cell lines (LCL) from unrelated individuals to SSRI drugs. Methods: The microRNAs miR-221, miR-222, and miR-151-a-3p, along with their target gene binding sites, were explored in silico using miRBase, TargetScan, microRNAviewer, and the UCSC Genome Browser. Luciferase reporter assays were conducted for demonstrating the direct functional regulation of ITGB3 and CHL1 expression by miR-221/222 and miR-151a-3p, respectively. A human LCL exhibiting low sensitivity to paroxetine was utilized for studying the phenotypic effect of CHL1 regulation by miR-151a-3p on SSRI response. Results: By showing direct regulation of CHL1 and ITGB3 by miR-151a-3p and miR-221/222, respectively, we link these microRNAs and genes with cellular SSRI sensitivity phenotypes. We report that miR-151a-3p increases cell sensitivity to paroxetine via down-regulating CHL1 expression. Conclusions: miR-151a-3p, miR-221/222 and their (here confirmed) respective target-genes, CHL1 and ITGB3, are implicated in SSRI responsiveness, and possibly in the clinical response to antidepressant drugs.

9.
Gene ; 606: 47-52, 2017 Mar 30.
Article in English | MEDLINE | ID: mdl-28089922

ABSTRACT

In this study, we report a family with X-linked recessive syndrome caused by mutated AMMECR1 and characterized by elliptocytosis with or without anemia, midface hypoplasia, proportionate short stature and hearing loss. Recently, mutations in AMMECR1 were reported in two maternal half-brothers, presenting with nephrocalcinosis, midface hypoplasia and, in one of the siblings, deafness and elliptocytosis. AMMECR1 gene is localized in the critical region of contiguous deletion syndrome on Xq22.3 implicated in Alport syndrome, mental retardation, midface hypoplasia, and elliptocytosis (AMME complex). Interestingly, alternative splicing of exon 2, the same exon harboring the truncating mutation, was observed in the proband and in his unaffected mother. Alternative splicing of this exon is predicted to lead to an in-frame deletion. We provide further evidence that mutated AMMECR1 gene is responsible for this clinically recognizable X-linked condition with variable expressivity.


Subject(s)
Craniofacial Abnormalities/genetics , Elliptocytosis, Hereditary/genetics , Genetic Diseases, X-Linked/genetics , Intellectual Disability/genetics , Nephritis, Hereditary/genetics , Proteins/genetics , Craniofacial Abnormalities/diagnosis , Craniofacial Abnormalities/pathology , Craniofacial Abnormalities/physiopathology , DNA Mutational Analysis , Elliptocytosis, Hereditary/diagnosis , Elliptocytosis, Hereditary/pathology , Elliptocytosis, Hereditary/physiopathology , Gene Deletion , Genetic Diseases, X-Linked/diagnosis , Genetic Diseases, X-Linked/pathology , Genetic Diseases, X-Linked/physiopathology , Humans , Intellectual Disability/diagnosis , Intellectual Disability/pathology , Intellectual Disability/physiopathology , Male , Nephritis, Hereditary/diagnosis , Nephritis, Hereditary/pathology , Nephritis, Hereditary/physiopathology , Proteins/chemistry , Proteins/metabolism
10.
J Immunol ; 196(12): 5156-65, 2016 06 15.
Article in English | MEDLINE | ID: mdl-27183614

ABSTRACT

The impact of microRNAs (miRNAs) known to regulate numerous biologic processes on complement-dependent cytotoxicity (CDC) was investigated in K562 cells. The C5b-9 complex is the executioner of CDC. Cells protect themselves from CDC by C5b-9 elimination, a process involving the mitochondrial chaperone mortalin/GRP75. Potential miR-200 (b and c) and miR-217 regulatory sites were identified in mortalin mRNA. Overexpression of miR-200b/c or miR-217 lowered the expression of mortalin mRNA. miRNA inhibitors for miR-200b, miR-200c, or miR-217 enhanced mortalin mRNA level. Unexpectedly, these miRNA modulators had no significant effect on mortalin protein level. Metabolic labeling analysis demonstrated that, to compensate for reduction in mortalin mRNA level, the cells increased the rate of synthesis of mortalin protein. Cells overexpressing miR-200b/c or miR-217 showed reduced sensitivity to CDC, whereas inhibition of miR-200c and miR-217 enhanced cell death. miR-200b/c overexpression reduced C5b-9 binding and enhanced its release from the cells and promoted mortalin relocation to the plasma membrane. Inhibition of miR-200 (b and c) and miR-217 had no effect on the expression level of the membrane complement-regulatory proteins CD46, CD55, and CD59. However, overexpression of miR-200b/c or miR-217 enhanced expression of CD46 and CD55 (not of CD59). Overall, the data demonstrate miRNA regulation of cell sensitivity to CDC. We identified miR-200b, miR-200c, and miR-217 as regulators of mortalin and, perhaps indirectly, of CD46 and CD55. Cell exposure to a sublytic dose of complement was shown to increase expression of miR-200 (b and c), suggesting that complement C5b-9 exerts a feedback-regulatory effect on these miRNAs.


Subject(s)
Complement Membrane Attack Complex/metabolism , Cytotoxicity, Immunologic , Gene Expression Regulation , HSP70 Heat-Shock Proteins/genetics , MicroRNAs/genetics , CD55 Antigens/genetics , CD59 Antigens/genetics , Cell Line, Tumor , Complement Membrane Attack Complex/immunology , Complement System Proteins/immunology , Humans , K562 Cells , Membrane Cofactor Protein/genetics , RNA, Messenger
11.
PLoS One ; 11(1): e0146236, 2016.
Article in English | MEDLINE | ID: mdl-26730965

ABSTRACT

Stress research has progressively become more integrative in nature, seeking to unfold crucial relations between the different phenotypic levels of stress manifestations. This study sought to unravel stress-induced variations in expression of human microRNAs sampled in peripheral blood mononuclear cells and further assess their relationship with neuronal and psychological indices. We obtained blood samples from 49 healthy male participants before and three hours after performing a social stress task, while undergoing functional magnetic resonance imaging (fMRI). A seed-based functional connectivity (FC) analysis was conducted for the ventro-medial prefrontal cortex (vmPFC), a key area of stress regulation. Out of hundreds of microRNAs, a specific increase was identified in microRNA-29c (miR-29c) expression, corresponding with both the experience of sustained stress via self-reports, and alterations in vmPFC functional connectivity. Explicitly, miR-29c expression levels corresponded with both increased connectivity of the vmPFC with the anterior insula (aIns), and decreased connectivity of the vmPFC with the left dorso-lateral prefrontal cortex (dlPFC). Our findings further revealed that miR-29c mediates an indirect path linking enhanced vmPFC-aIns connectivity during stress with subsequent experiences of sustained stress. The correlative patterns of miR-29c expression and vmPFC FC, along with the mediating effects on subjective stress sustainment and the presumed localization of miR-29c in astrocytes, together point to an intriguing assumption; miR-29c may serve as a biomarker in the blood for stress-induced functional neural alterations reflecting regulatory processes. Such a multi-level model may hold the key for future personalized intervention in stress psychopathology.


Subject(s)
Brain/physiopathology , MicroRNAs/metabolism , Stress, Psychological/metabolism , Epigenesis, Genetic , Functional Neuroimaging , Humans , Magnetic Resonance Imaging , Male , MicroRNAs/genetics , Neural Pathways/physiopathology , Stress, Psychological/genetics , Stress, Psychological/physiopathology , Young Adult
12.
Exp Dermatol ; 24(8): 618-22, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25939713

ABSTRACT

Alopecia-neurological defects-endocrinopathy (ANE) syndrome is a rare inherited hair disorder, which was shown to result from decreased expression of the RNA-binding motif protein 28 (RBM28). In this study, we attempted to delineate the role of RBM28 in hair biology. First, we sought to obtain evidence for the direct involvement of RBM28 in hair growth. When RBM28 was downregulated in human hair follicle (HF) organ cultures, we observed catagen induction and HF growth arrest, indicating that RBM28 is necessary for normal hair growth. We also aimed at identifying molecular targets of RBM28. Given that an RBM28 homologue was recently found to regulate miRNA biogenesis in C. elegans and given the known pivotal importance of miRNAs for proper hair follicle development, we studied global miRNA expression profile in cells knocked down for RBM28. This analysis revealed that RBM28 controls the expression of miR-203. miR-203 was found to regulate in turn TP63, encoding the transcription factor p63, which is critical for hair morphogenesis. In conclusion, RBM28 contributes to HF growth regulation through modulation of miR-203 and p63 activity.


Subject(s)
Alopecia/metabolism , Endocrine System Diseases/metabolism , Gene Expression Regulation , Hair Follicle/metabolism , Intellectual Disability/metabolism , MicroRNAs/physiology , RNA-Binding Proteins/physiology , Transcription Factors/physiology , Tumor Suppressor Proteins/physiology , Alopecia/physiopathology , Cells, Cultured , Endocrine System Diseases/physiopathology , Genes, Reporter , Hair/growth & development , Hair Follicle/growth & development , Humans , Intellectual Disability/physiopathology , Keratinocytes/metabolism , Morphogenesis , Organ Culture Techniques , RNA Interference , RNA, Small Interfering/genetics , RNA-Binding Proteins/genetics , Transfection , Up-Regulation
13.
J Cell Sci ; 128(8): 1607-16, 2015 Apr 15.
Article in English | MEDLINE | ID: mdl-25717002

ABSTRACT

MicroRNAs (miRNAs) are short non-coding RNAs that play a central role in the regulation of gene expression by binding to target mRNAs. Several studies have revealed alterations in cellular miRNA profiles following HIV-1 infection, mostly for miRNAs involved in inhibiting viral infection. These miRNA expression modifications might also serve to block the innate HIV-1 inhibition mechanism. As a result, it is expected that during HIV-1 infection miRNAs target genes that hinder or prevent the progression of the HIV-1 replication cycle. One of the major sets of genes known to inhibit the progression of HIV-1 infection are cellular restriction factors. In this study, we identified a direct miRNA target gene that modulates viral spread in T-lymphocytes and HeLa-CCR5 cell lines. Following infection, let-7c, miR-34a or miR-124a were upregulated, and they targeted and downregulated p21 and TASK1 (also known as CDKN1A and KCNK3, respectively) cellular proteins. This eventually led to increased virion release and higher copy number of viral genome transcripts in infected cells. Conversely, by downregulating these miRNAs, we could suppress viral replication and spread. Our data suggest that HIV-1 exploits the host miRNA cellular systems in order to block the innate inhibition mechanism, allowing a more efficient infection process.


Subject(s)
Cyclin-Dependent Kinase Inhibitor p21/metabolism , HIV-1/physiology , MicroRNAs/metabolism , Nerve Tissue Proteins/metabolism , Potassium Channels, Tandem Pore Domain/metabolism , Base Sequence , Down-Regulation , Gene Expression Regulation , HEK293 Cells , HIV Infections/metabolism , Humans , Molecular Sequence Data , RNA, Messenger/genetics , T-Lymphocytes/virology , Up-Regulation , Virus Replication
14.
Genet Res (Camb) ; 95(2-3): 53-6, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23938110

ABSTRACT

A recent E-Rare workshop reviewed the ethical aspects of whole exome and whole genome-sequencing studies (WES and WGS, respectively) in rare diseases. Leveraging new genomic technologies, which output vast amounts of known and novel genetic variants, researchers are learning more about the genetic basis and mechanisms involved in rare diseases. In some cases, these findings are translated into diagnostic tools for the benefit of rare disease patients. Among the disclosed data, which can assist in treatment management, incidental findings await, bringing with them ethical concerns for the clinicians, researchers and patients.


Subject(s)
Ethics, Medical , Exome , Genome, Human , Sequence Analysis, DNA , Humans , Israel
SELECTION OF CITATIONS
SEARCH DETAIL
...