RESUMO
Interactions between the splicing machinery and RNA polymerase II increase protein-coding gene transcription. Similarly, exons and splicing signals of enhancer-generated long noncoding RNAs (elncRNAs) augment enhancer activity. However, elncRNAs are inefficiently spliced, suggesting that, compared with protein-coding genes, they contain qualitatively different exons with a limited ability to drive splicing. We show here that the inefficiently spliced first exons of elncRNAs as well as promoter-antisense long noncoding RNAs (pa-lncRNAs) in human and mouse cells trigger a transcription termination checkpoint that requires WDR82, an RNA polymerase II-binding protein, and its RNA-binding partner of previously unknown function, ZC3H4. We propose that the first exons of elncRNAs and pa-lncRNAs are an intrinsic component of a regulatory mechanism that, on the one hand, maximizes the activity of these cis-regulatory elements by recruiting the splicing machinery and, on the other, contains elements that suppress pervasive extragenic transcription.
Assuntos
Proteínas Cromossômicas não Histona/genética , Proteínas de Ligação a DNA/genética , RNA Polimerase II/ultraestrutura , RNA Longo não Codificante/genética , Transcrição Gênica , Processamento Alternativo/genética , Animais , Proteínas Cromossômicas não Histona/ultraestrutura , Proteínas de Ligação a DNA/ultraestrutura , Éxons/genética , Humanos , Camundongos , Regiões Promotoras Genéticas/genética , RNA Polimerase II/genética , Splicing de RNA/genética , RNA Antissenso/genética , RNA Antissenso/ultraestrutura , RNA Longo não Codificante/ultraestrutura , RNA Mensageiro/genética , Sequências Reguladoras de Ácido Nucleico/genéticaRESUMO
The regulation of genomic function is in part mediated through the physical organization and architecture of the nucleus. Disruption to nuclear organization and architecture is increasingly being recognized by its contribution to many diseases. The LINC complexes - protein structures traversing the nuclear envelope, that physically connect the nuclear interior, and hence the genome, to cytoplasmic cytoskeletal networks are an important component in the physical organization of the genome and its function. This connection, potentially allows for the constant detection of environmental mechanical stimuli, resulting in altered regulation of nuclear architecture and genome function, either directly or via the process of mechanotransduction. Here, we review the influences LINC complexes exert on genome functions and their impact on cellular/organismal health.
Assuntos
Genoma/genética , Mecanotransdução Celular/genética , Proteínas Nucleares/genética , RNA Longo não Codificante/genética , Núcleo Celular/genética , Núcleo Celular/ultraestrutura , Citoplasma/genética , Citoplasma/ultraestrutura , Citoesqueleto/genética , Citoesqueleto/ultraestrutura , Humanos , Membrana Nuclear/genética , Membrana Nuclear/ultraestrutura , Proteínas Nucleares/ultraestrutura , RNA Longo não Codificante/ultraestruturaRESUMO
Long non-coding RNAs (lncRNAs) are attracting widespread attention for their emerging regulatory, transcriptional, epigenetic, structural and various other functions. Comprehensive transcriptome analysis has revealed that retrotransposon elements (REs) are transcribed and enriched in lncRNA sequences. However, the functions of lncRNAs and the molecular roles of the embedded REs are largely unknown. The secondary and tertiary structures of lncRNAs and their embedded REs are likely to have essential functional roles, but experimental determination and reliable computational prediction of large RNA structures have been extremely challenging. We report here the nuclear magnetic resonance (NMR)-based secondary structure determination of the 167-nt inverted short interspersed nuclear element (SINE) B2, which is embedded in antisense Uchl1 lncRNA and upregulates the translation of sense Uchl1 mRNAs. By using NMR 'fingerprints' as a sensitive probe in the domain survey, we successfully divided the full-length inverted SINE B2 into minimal units made of two discrete structured domains and one dynamic domain without altering their original structures after careful boundary adjustments. This approach allowed us to identify a structured domain in nucleotides 31-119 of the inverted SINE B2. This approach will be applicable to determining the structures of other regulatory lncRNAs.
Assuntos
Conformação de Ácido Nucleico , RNA Longo não Codificante/ultraestrutura , Retroelementos/genética , Elementos Nucleotídeos Curtos e Dispersos/genética , Biologia Computacional , Humanos , Espectroscopia de Ressonância Magnética , RNA Antissenso/genética , RNA Antissenso/ultraestrutura , RNA Longo não Codificante/genética , Transcriptoma/genética , Ubiquitina Tiolesterase/genética , Ubiquitina Tiolesterase/ultraestruturaRESUMO
Long noncoding RNA molecules (lncRNAs) are estimated to account for the majority of eukaryotic genomic transcripts, and have been associated with multiple diseases in humans. However, our understanding of their structure-function relationships is scarce, with structural evidence coming mostly from indirect biochemical approaches or computational predictions. Here we describe direct visualization of the lncRNA HOTAIR (HOx Transcript AntIsense RNA) using atomic force microscopy (AFM) in nucleus-like conditions at 37°. Our observations reveal that HOTAIR has a discernible, although flexible, shape. Fast AFM scanning enabled the quantification of the motion of HOTAIR, and provided visual evidence of physical interactions with genomic DNA segments. Our report provides a biologically plausible description of the anatomy and intrinsic properties of HOTAIR, and presents a framework for studying the structural biology of lncRNAs.
Assuntos
DNA/ultraestrutura , Conformação de Ácido Nucleico , RNA Longo não Codificante/ultraestrutura , Apoptose/genética , DNA/química , DNA/genética , Humanos , Microscopia de Força Atômica , RNA Longo não Codificante/química , RNA Longo não Codificante/genética , Relação Estrutura-AtividadeRESUMO
Long noncoding RNAs (lncRNAs) have been identified in all eukaryotes and are most abundant in the human genome. However, the functional importance and mechanisms of action for human lncRNAs are largely unknown. Using comparative sequence, structural, and functional analyses, we characterize the evolution and molecular function of human lncRNA JPX. We find that human JPX and its mouse homolog, lncRNA Jpx, have deep divergence in their nucleotide sequences and RNA secondary structures. Despite such differences, both lncRNAs demonstrate robust binding to CTCF, a protein that is central to Jpx's role in X chromosome inactivation. In addition, our functional rescue experiment using Jpx-deletion mutant cells shows that human JPX can functionally complement the loss of Jpx in mouse embryonic stem cells. Our findings support a model for functional conservation of lncRNAs independent from sequence and structural divergence. This study provides mechanistic insight into the evolution of lncRNA function.
Assuntos
Fator de Ligação a CCCTC/genética , Evolução Molecular , RNA Longo não Codificante/genética , Inativação do Cromossomo X/genética , Animais , Genoma Humano/genética , Humanos , Camundongos , Conformação de Ácido Nucleico , RNA Longo não Codificante/ultraestruturaRESUMO
Long non-coding RNAs (lncRNAs) have important roles in various cellular processes. Since the last couple of years the involvements of lncRNAs during the onset of various diseases are coming to our knowledge. Though the name is long non-coding RNAs, some of them are able to code for proteins. Analyses of lncRNAs, viz., ENSG00000235387, ENSG00000262179, ENSG00000233101 and ENSG00000204272 elucidated that they could code for small proteins/peptides. The proteins/peptides encoded by these lncRNAs are referred to as SPAR, Minion/myomixer, HOXB-AS3 and NOBODY respectively. So far only the amino acid sequence details regarding these peptides are available. Interestingly, all the works pertaining to lncRNAs are linked to the genetic aspects and till date no structural studies are carried out. Therefore, in this present work, an attempt has been made to analyze the peptides encoded by lncRNAs from a structural point of view. Their three dimensional structural frameworks have been identified and from there the possible binding modes of the peptides have been predicted. Till date, this is the first report that is aimed at understanding the molecular mechanism of the functionalities of the peptides encoded by these lncRNAs. This study would therefore be beneficial to future drug development endeavors to come up with new therapeutic strategies against the diseases to which these peptides are linked.