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The role of nuclear organization in trans-splicing based expression of heat shock protein 90 in giardia lamblia

Iyer, V and Tushir, S and Verma, S and Majumdar, S and Gayen, S and Mishra, R and Tatu, U (2021) The role of nuclear organization in trans-splicing based expression of heat shock protein 90 in giardia lamblia. In: PLoS Neglected Tropical Diseases, 15 (9).

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Official URL: https://doi.org/10.1371/journal.pntd.0009810


Hsp90 gene of G. lamblia has a split nature comprising two ORFs separated by 777 kb on chromosome 5. The ORFs of the split gene on chromosome 5 undergo transcription to generate independent pre-mRNAs that join by a unique trans-splicing reaction that remains partially understood. The canonical cis-acting nucleotide elements such as 5’SS-GU, 3’SS-AG, polypyrimidine tract and branch point adenine are present in the independent pre-mRNAs and therefore trans-splicing of Hsp90 must be assisted by spliceosomes in vivo. Using an approach of RNA-protein pull down, we show that an RNA helicase selectively interacts with HspN pre-mRNA. Our experiments involving high resolution chromosome conformation capture technology as well as DNA FISH show that the trans-spliced genes of Giardia are in three-dimensional spatial proximity in the nucleus. Altogether our study provides a glimpse into the in vivo mechanisms involving protein factors as well as chromatin structure to facili-tate the unique inter-molecular post-transcriptional stitching of split genes in G. lamblia.

Item Type: Journal Article
Publication: PLoS Neglected Tropical Diseases
Publisher: Public Library of Science
Additional Information: The copyright for this article belongs to the Authors.
Keywords: dynein adenosine triphosphatase; heat shock protein 90; RNA binding protein; RNA helicase; RNA polymerase; RNA polymerase II; heat shock protein 90; RNA precursor, Article; chromatin structure; chromosome 3; DNA sequence; fluorescence imaging; gene control; Giardia intestinalis; immunoblotting; in situ hybridization; in vitro study; mass spectrometry; organization; protein degradation; protein expression; protein function; protein RNA binding; RNA pull-down; trans splicing; transcription regulation; genetics; Giardia intestinalis; human; metabolism; open reading frame; RNA splicing; spliceosome, Giardia lamblia; HSP90 Heat-Shock Proteins; Humans; Open Reading Frames; RNA Precursors; RNA Splicing; Spliceosomes; Trans-Splicing
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 15 May 2023 09:51
Last Modified: 15 May 2023 09:51
URI: https://eprints.iisc.ac.in/id/eprint/81646

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