Interaction study of DHX15 helicase and 3’ end of Mason-Pfizer monkey virus RNA in vitro

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22810%2F25%3A43931952" target="_blank" >RIV/60461373:22810/25:43931952 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/60461373:22330/25:43931952

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Interaction study of DHX15 helicase and 3’ end of Mason-Pfizer monkey virus RNA in vitro

Popis výsledku v původním jazyce

The DHX15 helicase plays an important role in various steps of cellular RNA metabolism. To regulate its activity and increase its specificity, DHX15 requires activation by proteins containing a glycine-rich sequence known as the G-patch motif. This motif was identified in Mason-Pfizer monkey virus (MPMV) as a part of the polyprotein precursors Gag-Pro and Gag-Pro-Pol. In the Gag-Pro precursor, containing structural proteins and viral protease (PR), the G-patch motif is C-terminally fused to PR. In the Gag-Pro-Pol precursor (with Pol comprising reverse transcriptase (RT) and integrase), the G-patch motif acts as a linker between PR and RT. Functionally, the G-patch motif has been shown to positively influence virus infectivity and RT activity.The secondary structure of MPMV G-patch motif, consisting of a short brace helix and a brace loop, resembles that of cellular G-patch proteins known to interact with DHX15 (e.g., NF-kappa-B-repressing factor), suggesting a shared mode of action. Consistent with this, DHX15 was identified in purified MPMV particles, but only in virions containing an intact wt G-patch motif, not in those with mutated or deleted versions. Using PAR-CLIP analysis (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation), a short region at the 3´end of the MPMV genome was identified as a potential DHX15 binding site. This region is located upstream of the constitutive transport element (CTE), an RNA element crucial for the export of unspliced viral gRNA from the nucleus. To verify this interaction, we analyzed the binding of DHX15 to the identified region using microscale thermophoresis (MST).A deeper understanding of the interaction between DHX15 and retroviral gRNA could refine the model of unspliced viral gRNA export from the nucleus and provide new insight into viral RNA metabolism during the retroviral life cycle.

Název v anglickém jazyce

Interaction study of DHX15 helicase and 3’ end of Mason-Pfizer monkey virus RNA in vitro

Popis výsledku anglicky

The DHX15 helicase plays an important role in various steps of cellular RNA metabolism. To regulate its activity and increase its specificity, DHX15 requires activation by proteins containing a glycine-rich sequence known as the G-patch motif. This motif was identified in Mason-Pfizer monkey virus (MPMV) as a part of the polyprotein precursors Gag-Pro and Gag-Pro-Pol. In the Gag-Pro precursor, containing structural proteins and viral protease (PR), the G-patch motif is C-terminally fused to PR. In the Gag-Pro-Pol precursor (with Pol comprising reverse transcriptase (RT) and integrase), the G-patch motif acts as a linker between PR and RT. Functionally, the G-patch motif has been shown to positively influence virus infectivity and RT activity.The secondary structure of MPMV G-patch motif, consisting of a short brace helix and a brace loop, resembles that of cellular G-patch proteins known to interact with DHX15 (e.g., NF-kappa-B-repressing factor), suggesting a shared mode of action. Consistent with this, DHX15 was identified in purified MPMV particles, but only in virions containing an intact wt G-patch motif, not in those with mutated or deleted versions. Using PAR-CLIP analysis (Photoactivatable Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation), a short region at the 3´end of the MPMV genome was identified as a potential DHX15 binding site. This region is located upstream of the constitutive transport element (CTE), an RNA element crucial for the export of unspliced viral gRNA from the nucleus. To verify this interaction, we analyzed the binding of DHX15 to the identified region using microscale thermophoresis (MST).A deeper understanding of the interaction between DHX15 and retroviral gRNA could refine the model of unspliced viral gRNA export from the nucleus and provide new insight into viral RNA metabolism during the retroviral life cycle.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10607 - Virology

Projekt

<a href="/cs/project/LX22NPO5103" target="_blank" >LX22NPO5103: Národní institut virologie a bakteriologie</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů