Visualization of single mRNAs reveals temporal association of proteins with microRNA-regulated mRNA
نویسندگان
چکیده
Although many proteins are known to function in microRNA (miRNA)-based translational repression, we lack a comprehensive understanding of temporal relationships between the mRNA, miRNA and their constituent proteins. To understand the dynamics of miRNA and protein interactions, we created a synthetic inducible miRNA system in mammalian cells. By visualizing single mRNAs and observing their co-localization with proteins over time, we produced a temporal association map of miRNA-associated factors. Argonaute2, Dcp1a, hedls and Rck co-localize with miRNA-regulated mRNA after 24 h of miRNA induction, and RNAi knockdown of any one of these proteins affected the co-localization of any of the other proteins with miRNA-regulated mRNA, demonstrating that these proteins could interact with each other in a complex. We identified Argonaute2 and hedls as proteins that co-localize and interact with miRNA-regulated mRNA, indicating that processing body components are involved in long-term storage of miRNA-regulated mRNA.
منابع مشابه
Control of antagonistic components of the hedgehog signaling pathway by microRNAs in Drosophila.
Hedgehog (Hh) signaling is critical for many developmental processes and for the genesis of diverse cancers. Hh signaling comprises a series of negative regulatory steps, from Hh reception to gene transcription output. We previously showed that stability of antagonistic regulatory proteins, including the coreceptor Smoothened (Smo), the kinesin-like Costal-2 (Cos2), and the kinase Fused (Fu), i...
متن کاملMicroRNA Networks in Mouse Lung Organogenesis
BACKGROUND MicroRNAs (miRNAs) are known to be important regulators of both organ development and tumorigenesis. MiRNA networks and their regulation of messenger RNA (mRNA) translation and protein expression in specific biological processes are poorly understood. METHODS We explored the dynamic regulation of miRNAs in mouse lung organogenesis. Comprehensive miRNA and mRNA profiling was perform...
متن کاملMicroRNAs: Critical Regulators of mRNA Traffic and Translational Control with Promising Biotech and Therapeutic Applications
Context:MicroRNAs (miRNAs) are a class of short, endogenously-initiated, non-coding RNAs that post-transcriptionally control gene expression via translational repression or mRNA turnover. MiRNAs have attracted much attention in recent years as they play critical roles in gene expression and are promising tools with many biotech and therapeutic applications. The molecular mechanisms und...
متن کاملطرح تحلیل تعدادی از mRNA های مادری مخصوص اووسیت در جنین تک سلولی موش
Introduction & Objective: During oogenesis, mRNA is actively transcribed and accumulated in the growing oocytes, and then the transcription stops. Transcription silencing will continue during early embryonic stages at least up to the time when the embryonic genome is activated. Thus the earliest stages of embryogenesis in mammals and other animal species are depending on stored maternal RNAs an...
متن کاملIdentification of microRNA-regulated pathways using an integration of microRNA-mRNA microarray and bioinformatics analysis in CD34+ cells of myelodysplastic syndromes
The effect of microRNA (miRNA) and targeted mRNA on signal transduction is not fully understood in myelodysplastic syndromes (MDS). Here, we tried to identify the miRNAs-regulated pathways through a combination of miRNA and mRNA microarray in CD34+ cells from MDS patients. We identified 34 differentially expressed miRNAs and 1783 mRNAs in MDS. 25 dysregulated miRNAs and 394 targeted mRNAs were ...
متن کامل