RNA-protein interaction mediating post-transcriptional regulation in the circadian system.
Methods in molecular biology (Clifton, N.J.)(2009)
Abstract
Post-transcriptional control makes an important contribution to shaping transcript profiles of circadianly regulated genes. In Arabidopsis thaliana, the clock-regulated glycine-rich RNA-binding protein ATGRP7 oscillates with a 24-h rhythm and transmits the rhythmicity generated by the central oscillator within the cell. ATGRP7 negatively auto-regulates its own expression at the post-transcriptional level. In response to an elevated protein level, a shift to a cryptic 5' splice site within the intron occurs, leading to an unproductively spliced transcript that rapidly vanishes due to its short half-life. This feedback regulation relies on direct binding of the RNA-binding protein to its own RNA. Here we describe the analysis of RNA-protein interaction in vitro employing recombinant RNA-binding protein and 32P-labelled in vitro transcripts or synthetic RNA oligoribonucleotides comprising the binding site under study.
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Key words
circadian system,rna-protein,post-transcriptional
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