In vivo Validation of Hsp90 Trans-splicing in Giardia lamblia: Highlighting the Role of Cis-elements
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F24%3A10479587" target="_blank" >RIV/00216208:11310/24:10479587 - isvavai.cz</a>
Result on the web
<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=-elBL8Hru0" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=-elBL8Hru0</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.jmb.2024.168440" target="_blank" >10.1016/j.jmb.2024.168440</a>
Alternative languages
Result language
angličtina
Original language name
In vivo Validation of Hsp90 Trans-splicing in Giardia lamblia: Highlighting the Role of Cis-elements
Original language description
Giardia lamblia causes giardiasis, one of the most common human infectious diseases globally. Previous studies from our lab have shown that hsp90 gene of Giardia is split into two halves, namely hspN and hspC. The independent pre-mRNAs of these split genes join by trans -splicing, producing a full-length Hsp90 (FlHsp90) mRNA. Genetic manipulation of the participating genes is necessary to understand the mechanism and significance of such trans -splicing based expression of Hsp90. In this study, we have performed transfection based exogenous expression of hspN and/or hspC in G. lamblia. We electroporated a plasmid containing the Avi-tagged hspN component of Hsp90 and examined its fate in G. lamblia. We show that the exogenously expressed hspN RNA gets trans -spliced to endogenously expressed hspC RNA, giving rise to a hybrid-FlHsp90. We highlight the importance of cis -elements in this trans -splicing reaction through mutational analysis. The episomal plasmid carrying deletions in the intronic region of hspN, showed inhibition of the trans -splicing reaction. Additionally, exogenous hspC RNA also followed the same fate as of exogenous hspN, while upon co-transfection with episomal hspN, they underwent trans -splicing with each other. Using eGFP as a test protein, we have shown that intronic sequences of hsp90 gene can guide trans -splicing mediated repair of any associated exonic sequences. Our study provides in vivo validation of Hsp90 trans -splicing, showing crucial role of cis -elements and importantly highlights the potential of hsp90 intronic sequences to function as a minimal splicing tool.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10600 - Biological sciences
Result continuities
Project
<a href="/en/project/GA20-25417S" target="_blank" >GA20-25417S: Biogenesis and function of pathogen specific organelles.</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2024
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Molecular Biology
ISSN
0022-2836
e-ISSN
1089-8638
Volume of the periodical
436
Issue of the periodical within the volume
4
Country of publishing house
US - UNITED STATES
Number of pages
12
Pages from-to
168440
UT code for WoS article
001168039500001
EID of the result in the Scopus database
2-s2.0-85182940732