RESUMEN
Within cells, soluble RNPs can switch states to coassemble and condense into liquid or solid bodies. Although these phase transitions have been reconstituted in vitro, for endogenous bodies the diversity of the components, the specificity of the interaction networks, and the function of the coassemblies remain to be characterized. Here, by developing a fluorescence-activated particle sorting (FAPS) method to purify cytosolic processing bodies (P-bodies) from human epithelial cells, we identified hundreds of proteins and thousands of mRNAs that structure a dense network of interactions, separating P-body from non-P-body RNPs. mRNAs segregating into P-bodies are translationally repressed, but not decayed, and this repression explains part of the poor genome-wide correlation between RNA and protein abundance. P-bodies condense thousands of mRNAs that strikingly encode regulatory processes. Thus, we uncovered how P-bodies, by condensing and segregating repressed mRNAs, provide a physical substrate for the coordinated regulation of posttranscriptional mRNA regulons.
Asunto(s)
Regulación de la Expresión Génica , Proteoma/genética , ARN Mensajero/genética , Regulón , Ribonucleoproteínas/genética , Fraccionamiento Celular , Citoplasma/metabolismo , Gránulos Citoplasmáticos/química , Gránulos Citoplasmáticos/metabolismo , Ontología de Genes , Células HEK293 , Células HeLa , Humanos , Anotación de Secuencia Molecular , Transición de Fase , Biosíntesis de Proteínas , Proteoma/metabolismo , Estabilidad del ARN , ARN Mensajero/metabolismo , Ribonucleoproteínas/metabolismoRESUMEN
An increasing number of genomic tracks such as DNA methylation, histone modifications or transcriptomes are being produced to annotate genomes with functional states. The comparison of such high dimensional vectors obtained under various experimental conditions requires the use of a distance or dissimilarity measure. Pearson, Cosine and $L_{p}$-norm distances are commonly used for both count and binary vectors. In this article, we highlight how enhancement methods such as the contrast increasing mutual proximity' (MP) or local scaling' improve common distance measures. We present a systematic approach to evaluate the performance of such enhanced distance measures in terms of separability of groups of experimental replicates to outline their effect. We show that the MP' applied on the various distance measures drastically increases performance. Depending on the type of epigenetic experiment, MP' coupled together with Pearson, Cosine, $L_1$, Yule or Jaccard distances proves to be highly efficient in discriminating epigenomic profiles.
Asunto(s)
Genómica , Algoritmos , Epigenómica , MicroARNs/genética , ARN Mensajero/genéticaRESUMEN
While many computational methods have been proposed for 3D chromosome reconstruction from chromosomal contact maps, these methods are rarely used for the interpretation of such experimental data, in particular Hi-C data. We posit that this is due to the lack of an easy-to-use implementation of the proposed algorithms, as well as to the important computational cost of most methods. We here give a detailed implementation of the fast ShRec3D algorithm. We provide a tutorial that will enable the reader to reconstruct 3D consensus structures for human chromosomes and to decorate these structures with chromatin epigenetic states. We use this methodology to show that the bivalent chromatin, including Polycomb-rich domains, is spatially segregated and located in between the active and the quiescent chromatin compartments.