Open AccessThe role of nuclear organization in trans-splicing based expression of heat shock protein 90 in Giardia lamblia
Author(s) -
Vinithra Iyer,
Sheetal Tushir,
Shreekant Verma,
Sudeshna Majumdar,
Srimonta Gayen,
Rakesh K. Mishra,
Utpal Tatu
Publication year - 2021
Publication title -
plos neglected tropical diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.99
H-Index - 135
eISSN - 1935-2735
pISSN - 1935-2727
DOI - 10.1371/journal.pntd.0009810
Subject(s) - orfs , biology , giardia lamblia , trans splicing , rna splicing , genetics , gene , chromatin , spliceosome , transcription (linguistics) , gene expression , rna , microbiology and biotechnology , open reading frame , linguistics , philosophy , peptide sequence
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 facilitate the unique inter-molecular post-transcriptional stitching of split genes in G . lamblia .