Iyer, V and Chettiar, ST and Grover, M and Rajyaguru, P and Nageshan, RK and Tatu, U (2019) Giardia lamblia Hsp90 pre-mRNAs undergo self-splicing to generate mature RNA in an in vitro trans-splicing reaction. In: FEBS Letters, 593 (4). pp. 433-442.
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Abstract
We have previously shown that the Heat Shock Protein 90 (Hsp90) gene in G. lamblia is expressed from two ORFs localized 777 kb apart. The pre-mRNAs transcribed from these ORFs are stitched by a trans-splicing mechanism. Here, we provide mechanistic details of this process by reconstituting the reaction using in vitro synthesized pre-mRNA substrates. Using RT-PCR, northern blot and nanostring technology, we demonstrate that the in vitro synthesized pre-mRNAs have the capability to self-splice in the absence of nuclear proteins. Inhibition of the trans-splicing reaction using a ssDNA oligo corresponding to a 26-nucleotide complementary sequence confirmed their role in juxtapositioning the pre-mRNA substrates during the self-splicing reaction. Our study provides the first example of a self catalysed, trans-splicing reaction in eukaryotes.
| Item Type: | Journal Article |
|---|---|
| Publication: | FEBS Letters |
| Publisher: | Wiley Blackwell |
| Additional Information: | The copyright for this article belongs to the Authors. |
| Keywords: | heat shock protein 90; magnesium ion; messenger RNA precursor; single stranded DNA; heat shock protein 90; protozoal protein; RNA precursor; single stranded DNA, Article; controlled study; DNA probe; DNA sequence; Giardia intestinalis; hsp90 gene; in vitro study; nonhuman; Northern blotting; nucleophilicity; priority journal; real time polymerase chain reaction; RNA splice site; RNA synthesis; trans splicing; transcription regulation; genetics; Giardia intestinalis; metabolism; nanotechnology; open reading frame, DNA, Single-Stranded; Giardia lamblia; HSP90 Heat-Shock Proteins; Nanotechnology; Open Reading Frames; Protozoan Proteins; RNA Precursors; Trans-Splicing |
| Department/Centre: | Division of Biological Sciences > Biochemistry |
| Date Deposited: | 28 Oct 2022 06:24 |
| Last Modified: | 28 Oct 2022 06:24 |
| URI: | https://eprints.iisc.ac.in/id/eprint/77733 |
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