Interaction of Mason-Pfizer Monkey Virus Matrix Protein with Calmodulin: Effects on the Late-Phase of Retroviral Life Cycle

Kód výsledku v IS VaVaI

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RIV/60461373:22810/25:43931948

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Interaction of Mason-Pfizer Monkey Virus Matrix Protein with Calmodulin: Effects on the Late-Phase of Retroviral Life Cycle

Popis výsledku v původním jazyce

Calmodulin (CaM), a ubiquitous cellular protein, plays a critical role in regulating various cellular processes through calcium-induced conformational changes. Its involvement in the life cycle of many viruses, including retroviruses, is well documented. In HIV-1-infected cells, CaM concentration is significantly elevated compared to uninfected cells. The structural polyprotein Gag of HIV-1 interacts with CaM via its N-terminal matrix (MA) domain, a region crucial for viral targeting to the plasma membrane (PM). This interaction induces a myristoyl (myr) switch in the MA protein, exposing its N-terminal myristoyl group, which facilitates membrane binding.We extended this investigation to the Mason-Pfizer monkey virus (M-PMV), a retrovirus with distinct late-phase life cycle mechanisms. HIV-1 assembles at the plasma membrane (PM), whereas M-PMV assembles in the cytoplasm. Our co-immunoprecipitation experiments successfully demonstrated the interaction of M-PMV Gag with HA-tagged human calmodulin in HEK 293T cell lysates. This interaction was confirmed to be Ca²⁺-dependent in both in vivo and in vitro settings.Further characterization of the interaction was conducted using microscale thermophoresis, protein cross-linking coupled with mass spectrometry, and liposome binding assay. Additionally, MA mutants were engineered to assess their impact on the presence and parameters of the interaction. Our findings provide valuable insights, suggesting a potentially significant role for the MA-CaM interaction in regulating key steps of the late phase of the retroviral life cycle.

Název v anglickém jazyce

Interaction of Mason-Pfizer Monkey Virus Matrix Protein with Calmodulin: Effects on the Late-Phase of Retroviral Life Cycle

Popis výsledku anglicky

Calmodulin (CaM), a ubiquitous cellular protein, plays a critical role in regulating various cellular processes through calcium-induced conformational changes. Its involvement in the life cycle of many viruses, including retroviruses, is well documented. In HIV-1-infected cells, CaM concentration is significantly elevated compared to uninfected cells. The structural polyprotein Gag of HIV-1 interacts with CaM via its N-terminal matrix (MA) domain, a region crucial for viral targeting to the plasma membrane (PM). This interaction induces a myristoyl (myr) switch in the MA protein, exposing its N-terminal myristoyl group, which facilitates membrane binding.We extended this investigation to the Mason-Pfizer monkey virus (M-PMV), a retrovirus with distinct late-phase life cycle mechanisms. HIV-1 assembles at the plasma membrane (PM), whereas M-PMV assembles in the cytoplasm. Our co-immunoprecipitation experiments successfully demonstrated the interaction of M-PMV Gag with HA-tagged human calmodulin in HEK 293T cell lysates. This interaction was confirmed to be Ca²⁺-dependent in both in vivo and in vitro settings.Further characterization of the interaction was conducted using microscale thermophoresis, protein cross-linking coupled with mass spectrometry, and liposome binding assay. Additionally, MA mutants were engineered to assess their impact on the presence and parameters of the interaction. Our findings provide valuable insights, suggesting a potentially significant role for the MA-CaM interaction in regulating key steps of the late phase of the retroviral life cycle.

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