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1.
Proc Natl Acad Sci U S A ; 117(44): 27346-27353, 2020 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-33077600

RESUMO

A key functional event in eukaryotic gene activation is the formation of dynamic protein-protein interaction networks between transcriptional activators and transcriptional coactivators. Seemingly incongruent with the tight regulation of transcription, many biochemical and biophysical studies suggest that activators use nonspecific hydrophobic and/or electrostatic interactions to bind to coactivators, with few if any specific contacts. Here a mechanistic dissection of a set of representative dynamic activator•coactivator complexes, comprised of the ETV/PEA3 family of activators and the coactivator Med25, reveals a different molecular recognition model. The data demonstrate that small sequence variations within an activator family significantly redistribute the conformational ensemble of the complex while not affecting overall affinity, and distal residues within the activator-not often considered as contributing to binding-play a key role in mediating conformational redistribution. The ETV/PEA3•Med25 ensembles are directed by specific contacts between the disordered activator and the Med25 interface, which is facilitated by structural shifts of the coactivator binding surface. Taken together, these data highlight the critical role coactivator plasticity plays in recognition of disordered activators and indicate that molecular recognition models of disordered proteins must consider the ability of the binding partners to mediate specificity.


Assuntos
Fatores de Transcrição/metabolismo , Ativação Transcricional/genética , Sequência de Aminoácidos/genética , Humanos , Complexo Mediador/genética , Complexo Mediador/metabolismo , Modelos Moleculares , Ligação Proteica/genética , Domínios e Motivos de Interação entre Proteínas/genética , Ativação Transcricional/fisiologia
2.
Nucleic Acids Res ; 46(16): 8025-8037, 2018 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-30102391

RESUMO

Over the past decade, the RNA revolution has revealed thousands of non-coding RNAs that are essential for cellular regulation and are misregulated in disease. While the development of methods and tools to study these RNAs has been challenging, the power and promise of small molecule chemical probes is increasingly recognized. To harness existing knowledge, we compiled a list of 116 ligands with reported activity against RNA targets in biological systems (R-BIND). In this survey, we examine the RNA targets, design and discovery strategies, and chemical probe characterization techniques of these ligands. We discuss the applicability of current tools to identify and evaluate RNA-targeted chemical probes, suggest criteria to assess the quality of RNA chemical probes and targets, and propose areas where new tools are particularly needed. We anticipate that this knowledge will expedite the discovery of RNA-targeted ligands and the next phase of the RNA revolution.


Assuntos
Descoberta de Drogas , RNA não Traduzido/química , RNA/química , Bibliotecas de Moléculas Pequenas/química , Fenômenos Químicos , Humanos , Ligantes , RNA/antagonistas & inibidores , RNA não Traduzido/antagonistas & inibidores , Bibliotecas de Moléculas Pequenas/uso terapêutico
3.
Angew Chem Int Ed Engl ; 57(40): 13242-13247, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30134013

RESUMO

Structural studies of the 3'-end of the oncogenic long non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) confirmed a unique triple-helix structure. This structure enables accumulation of the transcript, and high levels of MALAT1 are found in several cancers. Here, we synthesize a small molecule library based on an RNA-binding scaffold, diphenylfuran (DPF), screen it against a variety of nucleic acid constructs, and demonstrate for the first time that the MALAT1 triple helix can be selectively targeted with small molecules. Computational analysis revealed a trend between subunit positioning and composition on DPF shape and intramolecular interactions, which in turn generally correlated with selectivity and binding strengths. This work thus provides design strategies toward chemical probe development for the MALAT1 triple helix and suggests that comprehensive analyses of RNA-focused libraries can generate insights into selective RNA recognition.


Assuntos
RNA Longo não Codificante/metabolismo , RNA/química , Bibliotecas de Moléculas Pequenas/química , Teoria da Densidade Funcional , Furanos/química , Furanos/metabolismo , Humanos , Ligantes , Conformação de Ácido Nucleico , RNA/metabolismo , RNA Longo não Codificante/química , Bibliotecas de Moléculas Pequenas/metabolismo
4.
Angew Chem Int Ed Engl ; 56(43): 13498-13502, 2017 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-28810078

RESUMO

While a myriad non-coding RNAs are known to be essential in cellular processes and misregulated in diseases, the development of RNA-targeted small molecule probes has met with limited success. To elucidate the guiding principles for selective small molecule/RNA recognition, we analyzed cheminformatic and shape-based descriptors for 104 RNA-targeted ligands with demonstrated biological activity (RNA-targeted BIoactive ligaNd Database, R-BIND). We then compared R-BIND to both FDA-approved small molecule drugs and RNA ligands without reported bioactivity. Several striking trends emerged for bioactive RNA ligands, including: 1) Compliance to medicinal chemistry rules, 2) distinctive structural features, and 3) enrichment in rod-like shapes over others. This work provides unique insights that directly facilitate the selection and synthesis of RNA-targeted libraries with the goal of efficiently identifying selective small molecule ligands for therapeutically relevant RNAs.


Assuntos
Ligantes , RNA/química , Bibliotecas de Moléculas Pequenas/química , Bases de Dados de Compostos Químicos , Descoberta de Drogas , Análise de Componente Principal , RNA/metabolismo , Bibliotecas de Moléculas Pequenas/metabolismo
5.
Biopolymers ; 104(4): 213-46, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25787087

RESUMO

Flavin-dependent, lysine-specific protein demethylases (KDM1s) are a subfamily of amine oxidases that catalyze the selective posttranslational oxidative demethylation of methyllysine side chains within protein and peptide substrates. KDM1s participate in the widespread epigenetic regulation of both normal and disease state transcriptional programs. Their activities are central to various cellular functions, such as hematopoietic and neuronal differentiation, cancer proliferation and metastasis, and viral lytic replication and establishment of latency. Interestingly, KDM1s function as catalytic subunits within complexes with coregulatory molecules that modulate enzymatic activity of the demethylases and coordinate their access to specific substrates at distinct sites within the cell and chromatin. Although several classes of KDM1-selective small molecule inhibitors have been recently developed, these pan-active site inhibition strategies lack the ability to selectively discriminate between KDM1 activity in specific, and occasionally opposing, functional contexts within these complexes. Here we review the discovery of this class of demethylases, their structures, chemical mechanisms, and specificity. Additionally, we review inhibition of this class of enzymes as well as emerging interactions with coregulatory molecules that regulate demethylase activity in highly specific functional contexts of biological and potential therapeutic importance.


Assuntos
Inibidores Enzimáticos/química , Histona Desmetilases/antagonistas & inibidores , Histona Desmetilases/química , Histona Desmetilases/metabolismo , Processamento de Proteína Pós-Traducional , Animais , Diferenciação Celular , Células-Tronco Hematopoéticas/enzimologia , Células-Tronco Hematopoéticas/patologia , Histona Desmetilases/classificação , Humanos , Neoplasias/enzimologia , Neoplasias/patologia , Células-Tronco Neurais/enzimologia , Células-Tronco Neurais/patologia
7.
bioRxiv ; 2023 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-37577658

RESUMO

Small molecules have become increasingly recognized as invaluable tools to study RNA structure and function and to develop RNA-targeted therapeutics. To rationally design RNA-targeting ligands, a comprehensive understanding and explicit testing of small molecule properties that govern molecular recognition is crucial. To date, most studies have primarily evaluated properties of small molecules that bind RNA in vitro, with little to no assessment of properties that are distinct to selective and bioactive RNA-targeted ligands. Therefore, we curated an RNA-focused library, termed the Duke RNA-Targeted Library (DRTL), that was biased towards the physicochemical and structural properties of biologically active and non-ribosomal RNA-targeted small molecules. The DRTL represents one of the largest academic RNA-focused small molecule libraries curated to date with more than 800 small molecules. These ligands were selected using computational approaches that measure similarity to known bioactive RNA ligands and that diversify the molecules within this space. We evaluated DRTL binding in vitro to a panel of four RNAs using two optimized fluorescent indicator displacement assays, and we successfully identified multiple small molecule hits, including several novel scaffolds for RNA. The DRTL has and will continue to provide insights into biologically relevant RNA chemical space, such as the identification of additional RNA-privileged scaffolds and validation of RNA-privileged molecular features. Future DRTL screening will focus on expanding both the targets and assays used, and we welcome collaboration from the scientific community. We envision that the DRTL will be a valuable resource for the discovery of RNA-targeted chemical probes and therapeutic leads.

8.
ACS Chem Biol ; 14(12): 2691-2700, 2019 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-31589399

RESUMO

While the opportunities available for targeting RNA with small molecules have been widely appreciated, the challenges associated with achieving specific RNA recognition in biological systems have hindered progress and prevented many researchers from entering the field. To facilitate the discovery of RNA-targeted chemical probes and their subsequent applications, we curated the RNA-targeted BIoactive ligaNd Database (R-BIND). This collection contains an array of information on reported chemical probes that target non-rRNA and have biological activity, and analysis has led to the discovery of RNA-privileged properties. Herein, we developed an online platform to make this information freely available to the community, offering search options, a suite of tools for probe development, and an updated R-BIND data set with detailed experimental information for each probe. We repeated the previous cheminformatics analysis on the updated R-BIND list and found that the distinguishing physicochemical, structural, and spatial properties remained unchanged, despite an almost 50% increase in the database size. Further, we developed several user-friendly tools, including queries based on cheminformatic parameters, experimental details, functional groups, and substructures. In addition, a nearest neighbor algorithm can assess the similarity of user-uploaded molecules to R-BIND ligands. These tools and resources can be used to design small molecule libraries, optimize lead ligands, or select targets, probes, assays, and control experiments. Chemical probes are critical to the study and discovery of novel functions for RNA, and we expect this resource to greatly assist researchers in exploring and developing successful RNA-targeted probes.


Assuntos
Bases de Dados de Ácidos Nucleicos , Sondas RNA/química , RNA/química , Algoritmos , Ligantes , Interface Usuário-Computador
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