EFFECT OF MYRISTOYL SWITCH INDUCED CLEAVAGE OF MATRIX DOMAIN ON THE LATE PHASE OF MASON-PFIZER MONKEY VIRUS LIFE CYCLE

Kód výsledku v IS VaVaI

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Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

EFFECT OF MYRISTOYL SWITCH INDUCED CLEAVAGE OF MATRIX DOMAIN ON THE LATE PHASE OF MASON-PFIZER MONKEY VIRUS LIFE CYCLE

Popis výsledku v původním jazyce

Retroviral particles formed during the late phase of the viral life cycle, must undergo maturation to become infectious. In this stepwise process, the viral polyproteins Gag and Gag-Pol are cleaved at specific sites by a virus-encoded protease. For C-type retroviruses, including HIV, where particles assemble at the plasma membrane (PM) and then directly bud through the PM, the trigger mechanism for maturation could involve both interactions with the PM and particle assembly that enables the viral protease to dimerize into an active enzyme. In contrast to C-type, D-type retroviruses such as Mason-Pfizer monkey virus (M-PMV) assemble in the cytoplasm. Preassembled intracytoplasmic viral particles are transported to the plasma membrane in their unprocessed form, where budding and maturation take place. Therefore, maturation in D-type retroviruses must be tightly regulated, and protease dimerization cannot be the mechanism controlling this process.We previously demonstrated that the M-PMV protease-mediated in vitro cleavage of myristoylated matrix (MA) domain, located at the N-terminus of Gag, from the downstream phosphoprotein (PP) domain of Gag is induced by the interaction of MA with liposomes. We further showed that this cleavage was facilitated by a structural change of the cleavage site induced by the MA-liposome interaction triggered myristoyl switch.To study the effect of myristoyl switch-induced MA domain cleavage on viral maturation in vivo, vectors were prepared to produce noninfectious viral particles with amino acid substitutions affecting MA myristoylation, myristoyl switch and blocking MA cleavage from the rest of the Gag. We showed that beta-branched amino acids, commonly used to block HIV-1 protease activity, were ineffective at blocking MA cleavage in M-PMV. However, we identified other amino acid substitutions that were able to block MA cleavage. Non-infectious M-PMV viral particles with above mentioned amino acid substitutions were produced in the HEK293T cell line and the effect of the amino acid substitutions on viral particle maturation was evaluated.

Název v anglickém jazyce

EFFECT OF MYRISTOYL SWITCH INDUCED CLEAVAGE OF MATRIX DOMAIN ON THE LATE PHASE OF MASON-PFIZER MONKEY VIRUS LIFE CYCLE

Popis výsledku anglicky

Retroviral particles formed during the late phase of the viral life cycle, must undergo maturation to become infectious. In this stepwise process, the viral polyproteins Gag and Gag-Pol are cleaved at specific sites by a virus-encoded protease. For C-type retroviruses, including HIV, where particles assemble at the plasma membrane (PM) and then directly bud through the PM, the trigger mechanism for maturation could involve both interactions with the PM and particle assembly that enables the viral protease to dimerize into an active enzyme. In contrast to C-type, D-type retroviruses such as Mason-Pfizer monkey virus (M-PMV) assemble in the cytoplasm. Preassembled intracytoplasmic viral particles are transported to the plasma membrane in their unprocessed form, where budding and maturation take place. Therefore, maturation in D-type retroviruses must be tightly regulated, and protease dimerization cannot be the mechanism controlling this process.We previously demonstrated that the M-PMV protease-mediated in vitro cleavage of myristoylated matrix (MA) domain, located at the N-terminus of Gag, from the downstream phosphoprotein (PP) domain of Gag is induced by the interaction of MA with liposomes. We further showed that this cleavage was facilitated by a structural change of the cleavage site induced by the MA-liposome interaction triggered myristoyl switch.To study the effect of myristoyl switch-induced MA domain cleavage on viral maturation in vivo, vectors were prepared to produce noninfectious viral particles with amino acid substitutions affecting MA myristoylation, myristoyl switch and blocking MA cleavage from the rest of the Gag. We showed that beta-branched amino acids, commonly used to block HIV-1 protease activity, were ineffective at blocking MA cleavage in M-PMV. However, we identified other amino acid substitutions that were able to block MA cleavage. Non-infectious M-PMV viral particles with above mentioned amino acid substitutions were produced in the HEK293T cell line and the effect of the amino acid substitutions on viral particle maturation was evaluated.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10607 - Virology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

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