SPATIOTEMPORAL STUDY OF MORPHO- FUNCTIONAL MODIFICATIONS ON CELL NUCLEUS DURING AFRICAN SWINE FEVER VIRUS EARLY PHASE OF INFECTION

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Spatiotemporal study of morpho-functional modifications on cell nucleus during African swine fever virus infection

Publication . Simões, Margarida Pires; Ferreira, Fernando António da Costa; Martins, Carlos Manuel Lopes Vieira

Studies on virus-host interactions are decisive to enhance our understanding on how African swine fever virus (ASFV) subverts cellular mechanisms, and also to better characterize host nucleus changes enabling this infection. Immunofluorescence studies and immunoblotting analysis of ASFV-infected cells, allowed us to identify the Ataxia telangiectasia mutated and Rad3-related (ATR) pathway as the specific DNA damage response (DDR) mechanism activated by ASFV infection. Additionally, the use of ATR kinase-dead cells confirmed that ATR has an essential role for the infection success. The viral intranuclear replication was then pursued using BrdU-pulse experiments, supported on previous reports about ASFV genome presence inside the host nucleus and the proven ATR activation. BrdU-labelled DNA molecules confirmed the active viral replication at early infection times, exclusively within the cell nucleus. Related spatial and morphological nuclear changes during ASFV infection were further addressed, particularly on subnuclear domains and host chromatin epigenetic signatures. Promyelocytic leukaemia nuclear bodies (PML-NBs), nuclear speckles and Cajal bodies displayed major alterations, accompanied by a repressive nuclear environment. PML knockdown revealed an essential proviral activity for ASFV successful infection. Herein, suggestions on how this work may help in the development of therapeutic strategies against ASFV infections can be found.

2015-09Tese de doutoramento

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Entidade financiadora

Fundação para a Ciência e a Tecnologia

Programa de financiamento

PIDDAC

Número da atribuição

SFRH/BD/65532/2009