High-throughput sequencing methods to study neuronal RNA-protein interactions.
UV-cross-linking and RNase protection, combined with high-throughput sequencing, have provided global maps of RNA sites bound by individual proteins or ribosomes. Using a stringent purification protocol, UV-CLIP (UV-cross-linking and immunoprecipitation) was able to identify intronic and exonic sites bound by splicing regulators in mouse brain tissue. Ribosome profiling has been used to quantify ribosome density on budding yeast mRNAs under different environmental conditions. Post-transcriptional regulation in neurons requires high spatial and temporal precision, as is evident from the role of localized translational control in synaptic plasticity. It remains to be seen if the high-throughput methods can be applied quantitatively to study the dynamics of RNP (ribonucleoprotein) remodelling in specific neuronal populations during the neurodegenerative process. It is certain, however, that applications of new biochemical techniques followed by high-throughput sequencing will continue to provide important insights into the mechanisms of neuronal post-transcriptional regulation.
Ule
Jernej
J
MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, UK. jule@mrc-lmb.cam.ac.uk
D000818:Animals; D001483:Base Sequence; D005091:Exons; D005786:Gene Expression Regulation; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000502:physiology*; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger