Abstract
The Regulation of Trypanosome Gene Expression by RNA-Binding Proteins
Highlights
Use a specific RNA as a ligand to purify proteins that bind to it3. Use a gel shift assay with a chosen RNA to monitor purification of a binding protein 4. Cross-link protein to a reporter RNA in vivo, then use that RNA to pull out the RNA-protein complex [31,32]
The genome organization of trypanosomes, leishmanias, and related kinetoplastid organisms is highly unusual: Each RNA polymerase II promoter precedes multiple open reading frames, and the primary transcript is cut into individual mRNAs by 59 trans splicing and 39 polyadenylation [1]
During translation the 59 cap is bound by initiation factors [3,4]; it is expected that, as in other eukaryotes, these interact with PABP, increasing translation efficiency and shielding the mRNA from degradation
Summary
3. Use a gel shift assay with a chosen RNA to monitor purification of a binding protein 4. Cross-link protein to a reporter RNA in vivo, then use that RNA to pull out the RNA-protein complex [31,32]
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