The Experts below are selected from a list of 129 Experts worldwide ranked by ideXlab platform
Kalpana White - One of the best experts on this subject based on the ideXlab platform.
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the third rna recognition motif of drosophila ELAV Protein has a role in multimerization
Nucleic Acids Research, 2008Co-Authors: Gakuta Toba, Kalpana WhiteAbstract:ELAV is a neuron-specific RNA-binding Protein in Drosophila that is required for development and maintenance of neurons. ELAV regulates alternative splicing of Neuroglian and erect wing (ewg) transcripts, and has been shown to form a multimeric complex on the last ewg intron. The Protein has three RNA recognition motifs (RRM1, 2 and 3) with a hinge region between RRM2 and 3. In this study, we used the yeast two-hybrid system to determine the multimerization domain of ELAV. Using deletion constructs, we mapped an interaction activity to a region containing most of RRM3. We found three conserved short sequences in RRM3 that were essential for the interaction, and also sufficient to give the interaction activity to RRM2 when introduced into it. In our in vivo functional assay, a mutation in one of the three sequences showed reduced activity in splicing regulation, underlining the functional importance of multimerization. However, RRM2 with the three RRM3 interaction sequences did not function as RRM3 in vivo, which suggested that multimerization is not the only function of RRM3. Our results are consistent with a model in which RRM3 serves as a bi-functional domain that interacts with both RNA and Protein.
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Neural specificity of ELAV expression: defining a Drosophila promoter for directing expression to the nervous system.
Journal of Neurochemistry, 2002Co-Authors: Kalpana WhiteAbstract:: The Drosophila melanogaster vital gene, embryonic lethal abnormal visual system (ELAV), is required for the postdeterminative development of the nervous system. Its gene product encodes an RNA binding Protein that was found to be expressed in all neurons right after their birth. This specific, ubiquitous, and continuous pattern of neural expression has led to the increasingly popular use of ELAV Protein as a neural-specific marker. To understand the molecular basis of this neural-specific expression, we have defined and analyzed the structure of the ELAV promoter. Cis-acting sequences important for conferring the neural specificity of ELAV expression were identified by analyzing the reporter gene expression in transformants carrying different ELAV-beta-galactosidase fusion genes. This analysis delimits a 333-bp region (-92 to +241) that is necessary for specifying the ELAV pattern of nervous system expression. A 3.5-kb promoter fragment encompassing this region was designed for targeting gene expression specifically to the nervous system and would be a useful tool for the analysis of nervous system function.
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Function of RRM domains of Drosophila melanogaster ELAV: Rnp1 mutations and rrm domain replacements with ELAV family Proteins and SXL.
Genetics, 2000Co-Authors: Michael J. Lisbin, Marshall Gordon, Yvonne M. Yannoni, Kalpana WhiteAbstract:Members of the ELAV family of Proteins contain three RNA recognition motifs (RRMs), which are highly conserved. ELAV, a Drosophila melanogaster member of this family, provides a vital function and exhibits a predominantly nuclear localization. To investigate if the RNA-binding property of each of the ELAV RRMs is required for ELAV's in vivo function, amino acid residues critical in RNA binding for each RRM were individually mutated. A stringent genetic complementation test revealed that when the mutant Protein was the sole source of ELAV, RNA-binding ability of each RRM was essential to ELAV function. To assess the degree to which each domain was specific for ELAV function and which domains perhaps performed a function common to related ELAV Proteins, we substituted an ELAV RRM with the corresponding RRM from RBP9, the D. melanogaster Protein most homologous to ELAV; HuD, a human ELAV family Protein; and SXL, which, although evolutionarily related, is not an ELAV family member. This analysis revealed that RRM3 replacements were fully functional, but RRM1 and RRM2 replacements were largely nonfunctional. Under less stringent conditions RRM1 and RRM2 replacements from SXL and RRM1 replacement from RBP9 were able to provide supplemental function in the presence of a mutant hypomorphic ELAV Protein.
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the neuron enriched splicing pattern of drosophila erect wing is dependent on the presence of ELAV Protein
Molecular and Cellular Biology, 2000Co-Authors: Sandhya P Koushika, Matthias Soller, Kalpana WhiteAbstract:Although the Drosophila melanogaster erect wing (ewg) gene is broadly transcribed in adults, an unusual posttranscriptional regulation involving alternative and inefficient splicing generates a 116-kDa EWG Protein in neurons, while Protein expression elsewhere or of other isoforms is below detection at this stage. This posttranscriptional control is important, as broad expression of EWG can be lethal. In this paper, we show that ELAV, a neuron-specific RNA binding Protein, is necessary to regulate EWG Protein expression in ELAV-null eye imaginal disc clones and that ELAV is sufficient for EWG expression in wing disc imaginal tissue after ectopic expression. Further, analysis of EWG expression elicited from intron-containing genomic transgenes and cDNA minitransgenes in ELAV-deficient eye discs shows that this regulation is dependent on the presence of ewg introns. Analyses of the ewg splicing patterns in wild-type and ELAV-deficient eye imaginal discs and in wild-type and ectopic ELAV-expressing wing imaginal discs, show that certain neuronal splice isoforms correspond to ELAV levels. The data presented in this paper are consistent with a mechanism in which ELAV increases the splicing efficiency of ewg transcripts in alternatively spliced regions rather than with a mechanism in which stability of specific splice forms is enhanced by ELAV. Additionally, we report that ELAV promotes a neuron-enriched splice isoform of Drosophila armadillo transcript. ELAV, however, is not involved in all neuron-enriched splice events.
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Domain necessary for Drosophila ELAV nuclear localization: function requires nuclear ELAV.
Journal of Cell Science, 1999Co-Authors: Yvonne M. Yannoni, Kalpana WhiteAbstract:The neuron specific Drosophila ELAV Protein belongs to the ELAV family of RNA binding Proteins which are characterized by three highly conserved RNA recognition motifs, an N-terminal domain, and a hinge region between the second and third RNA recognition motifs. Despite their highly conserved RNA recognition motifs the ELAV family members are a group of Proteins with diverse posttranscriptional functions including splicing regulation, mRNA stability and translatability and have a variety of subcellular localizations. The role of the ELAV hinge in localization and function was examined using transgenes encoding ELAV hinge deletions, in vivo. Subcellular localization of the hinge mutant Proteins revealed that residues between amino acids 333–374 are necessary for nuclear localization. This delineated sequence has no significant homology to classical nuclear localization sequences, but it is similar to the recently characterized nucleocytoplasmic shuttling sequence, the HNS, from a human ELAV family member, HuR. This defined sequence, however, was insufficient for nuclear localization as tested using hinge-GFP fusion Proteins. Functional assays revealed that mutant Proteins that fail to localize to the nucleus are unable to provide ELAV vital function, but their function is significantly restored when translocated into the nucleus by a heterologous nuclear localization sequence tag.
Simona Collina - One of the best experts on this subject based on the ideXlab platform.
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compounds interfering with embryonic lethal abnormal vision ELAV Protein rna complexes an avenue for discovering new drugs
Journal of Medicinal Chemistry, 2017Co-Authors: Rita Nasti, Daniela Rossi, Marialaura Amadio, Alessia Pascale, Yagiz M Unver, Anna K H Hirsch, Simona CollinaAbstract:RNA-binding Proteins play a key role in post-transcriptional processes. Among these Proteins, embryonic lethal abnormal vision (ELAV) Proteins are among the best described. ELAV Proteins predominantly act as positive regulators of gene expression, and their dysregulation is involved in several pathologies, such as cancer, inflammation, and neurodegenerative diseases. Only a few structurally unrelated compounds interfering with ELAV Protein–mRNA complexes have been identified by applying high-throughput screening approaches. Considering the structural diversity of the compounds discovered so far and the different techniques employed for screening their ability to interfere with ELAV Protein–mRNA complexes, drawing conclusions from structure–activity relationships remains a challenge. We performed docking studies to understand the interactions of compounds reported over the past decade to be inhibitors of ELAV Proteins and to evaluate the potential of computer-aided drug design to target this family of prot...
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Compounds Interfering with Embryonic Lethal Abnormal Vision (ELAV) Protein–RNA Complexes: An Avenue for Discovering New Drugs
Journal of Medicinal Chemistry, 2017Co-Authors: Rita Nasti, Daniela Rossi, Marialaura Amadio, Alessia Pascale, Anna K H Hirsch, M. Yagiz Unver, Simona CollinaAbstract:RNA-binding Proteins play a key role in post-transcriptional processes. Among these Proteins, embryonic lethal abnormal vision (ELAV) Proteins are among the best described. ELAV Proteins predominantly act as positive regulators of gene expression, and their dysregulation is involved in several pathologies, such as cancer, inflammation, and neurodegenerative diseases. Only a few structurally unrelated compounds interfering with ELAV Protein–mRNA complexes have been identified by applying high-throughput screening approaches. Considering the structural diversity of the compounds discovered so far and the different techniques employed for screening their ability to interfere with ELAV Protein–mRNA complexes, drawing conclusions from structure–activity relationships remains a challenge. We performed docking studies to understand the interactions of compounds reported over the past decade to be inhibitors of ELAV Proteins and to evaluate the potential of computer-aided drug design to target this family of prot...
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diffusion ordered spectroscopy and saturation transfer difference nmr spectroscopy studies of selective interactions between ELAV Protein fragments and an mrna target
European Journal of Organic Chemistry, 2014Co-Authors: Francesca Vasile, Daniela Rossi, Simona Collina, Donatella PotenzaAbstract:Recently, the crucial role of post-transcriptional processes in the control of gene expression opened a new fascinating route to the translation of basic mechanisms into clinical medicine. In this contest, Embryonic Lethal Abnormal Vision (ELAV) Proteins represent a family of well-characterized Proteins that bind mRNA molecules and affect their fate and, thus, the amount of corresponding Proteins. Interestingly, preliminary data bode well that the ELAV-mediated mRNA stabilization is a mechanism relevant to Alzheimer's disease. Herein, we report the analysis of four ELAV-derived peptides (P1–P4) belonging to the RNA recognition motif of ELAV Proteins in the presence of an mRNA fragment from tumor necrosis factor α. Advanced NMR techniques were used to gain insights into the interaction mechanism. Saturation transfer difference and diffusion-ordered NMR spectroscopy experiments showed that appropriate mixtures of peptides (and not each peptide individually) bind effectively to the biological target. The diffusion-ordered spectroscopy (DOSY) technique was applied here for the first time to the study of mRNA–peptides complexes. For each peptide, the 3D structure was also determined by NOE analysis supported by molecular dynamics simulations.
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Diffusion‐Ordered Spectroscopy and Saturation Transfer Difference NMR Spectroscopy Studies of Selective Interactions between ELAV Protein Fragments and an mRNA Target
European Journal of Organic Chemistry, 2014Co-Authors: Francesca Vasile, Daniela Rossi, Simona Collina, Donatella PotenzaAbstract:Recently, the crucial role of post-transcriptional processes in the control of gene expression opened a new fascinating route to the translation of basic mechanisms into clinical medicine. In this contest, Embryonic Lethal Abnormal Vision (ELAV) Proteins represent a family of well-characterized Proteins that bind mRNA molecules and affect their fate and, thus, the amount of corresponding Proteins. Interestingly, preliminary data bode well that the ELAV-mediated mRNA stabilization is a mechanism relevant to Alzheimer's disease. Herein, we report the analysis of four ELAV-derived peptides (P1–P4) belonging to the RNA recognition motif of ELAV Proteins in the presence of an mRNA fragment from tumor necrosis factor α. Advanced NMR techniques were used to gain insights into the interaction mechanism. Saturation transfer difference and diffusion-ordered NMR spectroscopy experiments showed that appropriate mixtures of peptides (and not each peptide individually) bind effectively to the biological target. The diffusion-ordered spectroscopy (DOSY) technique was applied here for the first time to the study of mRNA–peptides complexes. For each peptide, the 3D structure was also determined by NOE analysis supported by molecular dynamics simulations.
Donatella Potenza - One of the best experts on this subject based on the ideXlab platform.
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diffusion ordered spectroscopy and saturation transfer difference nmr spectroscopy studies of selective interactions between ELAV Protein fragments and an mrna target
European Journal of Organic Chemistry, 2014Co-Authors: Francesca Vasile, Daniela Rossi, Simona Collina, Donatella PotenzaAbstract:Recently, the crucial role of post-transcriptional processes in the control of gene expression opened a new fascinating route to the translation of basic mechanisms into clinical medicine. In this contest, Embryonic Lethal Abnormal Vision (ELAV) Proteins represent a family of well-characterized Proteins that bind mRNA molecules and affect their fate and, thus, the amount of corresponding Proteins. Interestingly, preliminary data bode well that the ELAV-mediated mRNA stabilization is a mechanism relevant to Alzheimer's disease. Herein, we report the analysis of four ELAV-derived peptides (P1–P4) belonging to the RNA recognition motif of ELAV Proteins in the presence of an mRNA fragment from tumor necrosis factor α. Advanced NMR techniques were used to gain insights into the interaction mechanism. Saturation transfer difference and diffusion-ordered NMR spectroscopy experiments showed that appropriate mixtures of peptides (and not each peptide individually) bind effectively to the biological target. The diffusion-ordered spectroscopy (DOSY) technique was applied here for the first time to the study of mRNA–peptides complexes. For each peptide, the 3D structure was also determined by NOE analysis supported by molecular dynamics simulations.
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Diffusion‐Ordered Spectroscopy and Saturation Transfer Difference NMR Spectroscopy Studies of Selective Interactions between ELAV Protein Fragments and an mRNA Target
European Journal of Organic Chemistry, 2014Co-Authors: Francesca Vasile, Daniela Rossi, Simona Collina, Donatella PotenzaAbstract:Recently, the crucial role of post-transcriptional processes in the control of gene expression opened a new fascinating route to the translation of basic mechanisms into clinical medicine. In this contest, Embryonic Lethal Abnormal Vision (ELAV) Proteins represent a family of well-characterized Proteins that bind mRNA molecules and affect their fate and, thus, the amount of corresponding Proteins. Interestingly, preliminary data bode well that the ELAV-mediated mRNA stabilization is a mechanism relevant to Alzheimer's disease. Herein, we report the analysis of four ELAV-derived peptides (P1–P4) belonging to the RNA recognition motif of ELAV Proteins in the presence of an mRNA fragment from tumor necrosis factor α. Advanced NMR techniques were used to gain insights into the interaction mechanism. Saturation transfer difference and diffusion-ordered NMR spectroscopy experiments showed that appropriate mixtures of peptides (and not each peptide individually) bind effectively to the biological target. The diffusion-ordered spectroscopy (DOSY) technique was applied here for the first time to the study of mRNA–peptides complexes. For each peptide, the 3D structure was also determined by NOE analysis supported by molecular dynamics simulations.
Alessia Pascale - One of the best experts on this subject based on the ideXlab platform.
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compounds interfering with embryonic lethal abnormal vision ELAV Protein rna complexes an avenue for discovering new drugs
Journal of Medicinal Chemistry, 2017Co-Authors: Rita Nasti, Daniela Rossi, Marialaura Amadio, Alessia Pascale, Yagiz M Unver, Anna K H Hirsch, Simona CollinaAbstract:RNA-binding Proteins play a key role in post-transcriptional processes. Among these Proteins, embryonic lethal abnormal vision (ELAV) Proteins are among the best described. ELAV Proteins predominantly act as positive regulators of gene expression, and their dysregulation is involved in several pathologies, such as cancer, inflammation, and neurodegenerative diseases. Only a few structurally unrelated compounds interfering with ELAV Protein–mRNA complexes have been identified by applying high-throughput screening approaches. Considering the structural diversity of the compounds discovered so far and the different techniques employed for screening their ability to interfere with ELAV Protein–mRNA complexes, drawing conclusions from structure–activity relationships remains a challenge. We performed docking studies to understand the interactions of compounds reported over the past decade to be inhibitors of ELAV Proteins and to evaluate the potential of computer-aided drug design to target this family of prot...
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Compounds Interfering with Embryonic Lethal Abnormal Vision (ELAV) Protein–RNA Complexes: An Avenue for Discovering New Drugs
Journal of Medicinal Chemistry, 2017Co-Authors: Rita Nasti, Daniela Rossi, Marialaura Amadio, Alessia Pascale, Anna K H Hirsch, M. Yagiz Unver, Simona CollinaAbstract:RNA-binding Proteins play a key role in post-transcriptional processes. Among these Proteins, embryonic lethal abnormal vision (ELAV) Proteins are among the best described. ELAV Proteins predominantly act as positive regulators of gene expression, and their dysregulation is involved in several pathologies, such as cancer, inflammation, and neurodegenerative diseases. Only a few structurally unrelated compounds interfering with ELAV Protein–mRNA complexes have been identified by applying high-throughput screening approaches. Considering the structural diversity of the compounds discovered so far and the different techniques employed for screening their ability to interfere with ELAV Protein–mRNA complexes, drawing conclusions from structure–activity relationships remains a challenge. We performed docking studies to understand the interactions of compounds reported over the past decade to be inhibitors of ELAV Proteins and to evaluate the potential of computer-aided drug design to target this family of prot...
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Autophagy stimulus affects different kinase pathways and promotes HuR Protein activation and SQSTM1/p62 Protein synthesis in ARPE-19 cells
Acta Ophthalmologica, 2015Co-Authors: Marialaura Amadio, Alessia Pascale, Stefano Govoni, Nicoletta Marchesi, Kai KaarnirantaAbstract:Purpose Age-related macular degeneration (AMD) pathogenesis is characterized by Protein degradation impairment in retinal pigment epithelial (RPE) cells. We previously found that the expression of autophagy receptor SQSTM1/p62 is positively regulated by the RNA-binding HuR/ELAV Protein. We investigated the effects of AICAR (autophagy inducer, 5-aminoimidazole-4- carboxamide-1-β-D-ribofuranoside) and MG-132 (proteasome inhibitor) co-treatment on HuR activation, p62 expression, and the kinases potentially involved. Methods ARPE-19 cells were treated with MG-132 (1 µM) and/or AICAR (2 mM) for increasing times (up to 2 h) and subjected to cell fractionation. SQSTM1/p62 mRNA and Protein levels were measured by qRT-PCR and Western blotting, respectively. HuR Protein levels and its phosphorylated status were evaluated by Western blotting. The effects of puromycin (1 mM, Protein synthesis inhibitor) and various kinase inhibitors were also tested. Results AICAR+MG-132 co-treatment for 2 h induces HuR Protein up-regulation, its cytoplasmic translocation and phosphorylation, as well as increased expression of p62 Protein, being the latter one blunted by puromycin. AICAR+MG-132 co-treatment affects various kinases with differential outcomes. Conclusions AICAR+MG-132 co-treatment leads to HuR activation and p62 Protein translation. Different Protein kinase pathways are likely involved in these events.
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Protein kinase c activation affects via the mrna binding hu antigen r ELAV Protein vascular endothelial growth factor expression in a pericytic endothelial coculture model
Molecular Vision, 2012Co-Authors: Marialaura Amadio, Cecilia Osera, Gabriella Lupo, Carla Motta, Filippo Drago, S Govoni, Alessia PascaleAbstract:Purpose: To explore whether, following direct contact, there is mutual influence between pericytes (PC) and endothelial cells (EC), and to establish whether Protein kinase C (PKC) activation, a condition associated with hyperglycemia, can affect, via the mRNA-binding Hu-antigen R (HuR)/ELAV Protein, the expression of vascular endothelial growth factor (VEGF). Methods: PC and EC were cultured separately or in direct contact (1:1 ratio), and exposed or not to phorbol esters, a PKC activator (100 nM for 15 min). Barrier integrity was evaluated by measuring endothelial electrical resistance and permeability to sodium fluorescein. Immunocytochemistry was performed to visualize EC and PC in coculture, and to evaluate phorbol 12-myristate-13-acetate (PMA)-induced HuR translocation. PKCβI/βII, HuR, and VEGF Protein content was measured with western blotting, VEGF secretion in cell culture medium was evaluated with enzyme-linked immunosorbent assay (ELISA), and quantification of VEGF mRNA was performed with real-time quantitative PCR. Results: In monocultures, VEGF mRNA/Protein basal levels were more elevated in PC than in EC. However, the basal expression of VEGF Protein, but not mRNA, in PC and EC was affected by culture conditions. In fact, physical contact with PC upregulated VEGF Protein levels in the EC, while VEGF was downregulated in PC cocultured with EC. In this last condition, PKCβII and HuR Protein basal levels were also decreased in monocultured PC. Moreover, in basal conditions, the amount of VEGF released from the coculture was higher than from the monocultures. Direct activation of PKCβ induced HuR translocation from the nuclear area to the cytoplasm, and increased the Protein levels of the kinase itself, HuR, and VEGF in PC and EC in both culture conditions. Concerning VEGF mRNA, PKC activation induced an increase in PKC levels only in monocultured EC and, conversely, a significant decrease in the same transcript amount in cocultured PC. PMA stimulus also led to a significant increase in VEGF secretion in coculture. Conclusions: When cocultured with PC, EC form a significantly tighter barrier than the endothelial monolayer. The physical contact leads to opposite changes in VEGF Protein levels in PC and EC. In particular, in basal conditions, cocultured PC seemed to downregulate their own expression of this proproliferating factor, as well as that of PKCβII and HuR, likely to maintain the 1:1 ratio with the cohabiting EC. In mono- and cocultured PC/EC, PKC direct activation led to a similar increase in PKCβI/βII, HuR, and VEGF Protein levels, changes that may also occur at early stages of diabetic retinopathy. The release of VEGF in the medium was favored by physical contact between PC and EC and was further increased by PMA exposure. In contrast with the effects on VEGF Protein, PKCβ activation induced modifications in VEGF mRNA content that are different in function of the cell type and the culture conditions. These findings suggest that the changes in the VEGF Protein and transcript observed in PC/EC can be ascribed to distinct and concomitant pathways. Further studies on this in vitro coculture model would be useful to better understand the PC/EC interaction in physiologic and pathological conditions. Diabetic retinopathy accounts for about 5% of cases of blindness worldwide, making diabetes the leading cause of vision loss in working-age adults. Although in some cases diabetic retinopathy signs are localized in the central macula and symptoms may be observed at early stages, the majority of patients with diabetic retinopathy are asymptomatic until the late stages of the disease [1], emphasizing the need to
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The PKCβ/HuR/VEGF pathway in diabetic retinopathy
Biochemical Pharmacology, 2010Co-Authors: Marialaura Amadio, Filippo Drago, Claudio Bucolo, Gm Leggio, Stefano Govoni, Alessia PascaleAbstract:We investigated whether the diabetes-related PKCβ activation affects VEGF expression through the mRNA-stabilizing human embryonic lethal abnormal vision (ELAV) Protein, HuR, in the retina of streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in rats by STZ injection. Retinal tissues were processed to detect PKCβI, PKCβII, VEGF and HuR contents, as well as HuR phosphorylation. Immunoprecipitation coupled to RT-PCR was employed to evaluate HuR binding to VEGF mRNA in RiboNucleoProteic (RNP) complexes. Statistical analysis was performed by ANOVA followed by an appropriate post hoc comparison test. Following experimental diabetes PKCβI and PKCβII levels were increased compared to sham; there was also a PKC-mediated phosphorylation/activation of HuR. These effects were blunted by the in vivo co-administration of a selective PKCβ inhibitor. A specific binding between the HuR Protein and the VEGF mRNA was also detected. The PKCβ/HuR activation was accompanied by enhanced VEGF Protein expression that was, again, blunted by the PKCβ inhibitor. These findings first demonstrate the activation, in the retina, of the PKCβ/HuR/VEGF pathway following experimental diabetes and disclose a new potential pharmacological target to counteract pathologies implicating VEGF deregulation, such as diabetic retinopathy.
Daniela Rossi - One of the best experts on this subject based on the ideXlab platform.
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compounds interfering with embryonic lethal abnormal vision ELAV Protein rna complexes an avenue for discovering new drugs
Journal of Medicinal Chemistry, 2017Co-Authors: Rita Nasti, Daniela Rossi, Marialaura Amadio, Alessia Pascale, Yagiz M Unver, Anna K H Hirsch, Simona CollinaAbstract:RNA-binding Proteins play a key role in post-transcriptional processes. Among these Proteins, embryonic lethal abnormal vision (ELAV) Proteins are among the best described. ELAV Proteins predominantly act as positive regulators of gene expression, and their dysregulation is involved in several pathologies, such as cancer, inflammation, and neurodegenerative diseases. Only a few structurally unrelated compounds interfering with ELAV Protein–mRNA complexes have been identified by applying high-throughput screening approaches. Considering the structural diversity of the compounds discovered so far and the different techniques employed for screening their ability to interfere with ELAV Protein–mRNA complexes, drawing conclusions from structure–activity relationships remains a challenge. We performed docking studies to understand the interactions of compounds reported over the past decade to be inhibitors of ELAV Proteins and to evaluate the potential of computer-aided drug design to target this family of prot...
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Compounds Interfering with Embryonic Lethal Abnormal Vision (ELAV) Protein–RNA Complexes: An Avenue for Discovering New Drugs
Journal of Medicinal Chemistry, 2017Co-Authors: Rita Nasti, Daniela Rossi, Marialaura Amadio, Alessia Pascale, Anna K H Hirsch, M. Yagiz Unver, Simona CollinaAbstract:RNA-binding Proteins play a key role in post-transcriptional processes. Among these Proteins, embryonic lethal abnormal vision (ELAV) Proteins are among the best described. ELAV Proteins predominantly act as positive regulators of gene expression, and their dysregulation is involved in several pathologies, such as cancer, inflammation, and neurodegenerative diseases. Only a few structurally unrelated compounds interfering with ELAV Protein–mRNA complexes have been identified by applying high-throughput screening approaches. Considering the structural diversity of the compounds discovered so far and the different techniques employed for screening their ability to interfere with ELAV Protein–mRNA complexes, drawing conclusions from structure–activity relationships remains a challenge. We performed docking studies to understand the interactions of compounds reported over the past decade to be inhibitors of ELAV Proteins and to evaluate the potential of computer-aided drug design to target this family of prot...
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diffusion ordered spectroscopy and saturation transfer difference nmr spectroscopy studies of selective interactions between ELAV Protein fragments and an mrna target
European Journal of Organic Chemistry, 2014Co-Authors: Francesca Vasile, Daniela Rossi, Simona Collina, Donatella PotenzaAbstract:Recently, the crucial role of post-transcriptional processes in the control of gene expression opened a new fascinating route to the translation of basic mechanisms into clinical medicine. In this contest, Embryonic Lethal Abnormal Vision (ELAV) Proteins represent a family of well-characterized Proteins that bind mRNA molecules and affect their fate and, thus, the amount of corresponding Proteins. Interestingly, preliminary data bode well that the ELAV-mediated mRNA stabilization is a mechanism relevant to Alzheimer's disease. Herein, we report the analysis of four ELAV-derived peptides (P1–P4) belonging to the RNA recognition motif of ELAV Proteins in the presence of an mRNA fragment from tumor necrosis factor α. Advanced NMR techniques were used to gain insights into the interaction mechanism. Saturation transfer difference and diffusion-ordered NMR spectroscopy experiments showed that appropriate mixtures of peptides (and not each peptide individually) bind effectively to the biological target. The diffusion-ordered spectroscopy (DOSY) technique was applied here for the first time to the study of mRNA–peptides complexes. For each peptide, the 3D structure was also determined by NOE analysis supported by molecular dynamics simulations.
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Diffusion‐Ordered Spectroscopy and Saturation Transfer Difference NMR Spectroscopy Studies of Selective Interactions between ELAV Protein Fragments and an mRNA Target
European Journal of Organic Chemistry, 2014Co-Authors: Francesca Vasile, Daniela Rossi, Simona Collina, Donatella PotenzaAbstract:Recently, the crucial role of post-transcriptional processes in the control of gene expression opened a new fascinating route to the translation of basic mechanisms into clinical medicine. In this contest, Embryonic Lethal Abnormal Vision (ELAV) Proteins represent a family of well-characterized Proteins that bind mRNA molecules and affect their fate and, thus, the amount of corresponding Proteins. Interestingly, preliminary data bode well that the ELAV-mediated mRNA stabilization is a mechanism relevant to Alzheimer's disease. Herein, we report the analysis of four ELAV-derived peptides (P1–P4) belonging to the RNA recognition motif of ELAV Proteins in the presence of an mRNA fragment from tumor necrosis factor α. Advanced NMR techniques were used to gain insights into the interaction mechanism. Saturation transfer difference and diffusion-ordered NMR spectroscopy experiments showed that appropriate mixtures of peptides (and not each peptide individually) bind effectively to the biological target. The diffusion-ordered spectroscopy (DOSY) technique was applied here for the first time to the study of mRNA–peptides complexes. For each peptide, the 3D structure was also determined by NOE analysis supported by molecular dynamics simulations.