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The combination of glutamate receptor antagonist MK-801 with tamoxifen and its ...
Ribeiro, Mariana P. C.; Nunes-Correia, Isabel; Santos, Armanda E.; Custódio, José B. A.Recent reports suggest that N-methyl-d-aspartate receptor (NMDAR) blockade by MK-801 decreases tumor growth. Thus, we investigated whether other ionotropic glutamate receptor (iGluR) antagonists were also able to modulate the proliferation of melanoma cells. On the other hand, the antiestrogen tamoxifen (TAM) decreases the proliferation of melanoma cells, and is included in combined therapies for melanoma. As t...
African swine fever virus p10 protein exhibits nuclear import capacity and accu...
Nunes-Correia, Isabel; Rodríguez, Javier M.; Eulálio, Ana; Carvalho, Ana L.; Citovsky, Vitaly; Simões, Sérgio; Faro, Carlos; Salas, María L.African swine fever virus (ASFV), a large enveloped DNA-containing virus, infects domestic and wild pigs, and multiplies in soft ticks, causing an economically relevant hemorrhagic disease. Evaluation of the nuclear import ability of ASFV p10 protein was the major purpose of the present work. Two approaches were used to determine if p10 protein is imported into the nucleus by an active process: a yeast-based nu...
African swine fever virus p37 structural protein is localized in nuclear foci c...
Eulálio, Ana; Nunes-Correia, Isabel; Salas, José; Salas, Maria L.; Simões, Sergio; Pedroso de Lima, Maria C.The replication of African swine fever virus DNA is initiated inside the nucleus of host cells, being followed by a longer cytoplasmic replication stage. In face of previous results demonstrating the nucleo-cytoplasmic shuttling activity of ASFV p37 structural protein when considered isolated from the virus infection, we performed a systematic analysis of the subcellular localization of p37 protein in ASFV-infe...
Nuclear export of African swine fever virus p37 protein occurs through two dist...
Eulálio, Ana; Nunes-Correia, Isabel; Carvalho, Ana Luísa; Faro, Carlos; Citovsky, Vitaly; Salas, José; Salas, Maria L.; Simões, SérgioNucleocytoplasmic shuttling activity of the African swine fever virus p37 protein, a major structural protein of this highly complex virus, has been recently reported. The systematic characterization of the nuclear export ability of this protein constituted the major purpose of the present study. We report that both the N- and C-terminal regions of p37 protein are actively exported from the nucleus to the cytop...
Caveolae as an additional route for influenza virus endocytosis in MDCK cells
Nunes-Correia, Isabel; Eulálio, Ana; Nir, Shlomo; Lima, Maria C. Pedroso deClathrin-mediated endocytosis has been described as the primary internalization pathway for many viruses, including the influenza virus. However, caveolae, an alternative clathrin-independent endocytotic pathway, has also been described as mediating the entry of some molecules, including viruses. To address the question of pathway selection by the influenza virus, we have investigated whether the virus is inter...
Fluorescent probes for monitoring virus fusion kinetics: comparative evaluation...
Nunes-Correia, Isabel; Eulálio, Ana; Nir, Shlomo; Düzgünes, Nejat; Ramalho-Santos, João; Lima, Maria C. Pedroso deFluorescence assays for viral membrane fusion employ lipidic probes whose kinetics of fluorescence dequenching should mimic the actual kinetics of membrane merging. We examined the fusion of influenza virus with CEM cells, erythrocyte ghosts or liposomes by monitoring the fluorescence dequenching of each one of the three probes, octadecylrhodamine B chloride (R18), N-(lissamine rhodamine B sulfonyl)phosphatidyl...
Interactions of Influenza Virus with Cultured Cells: Detailed Kinetic Modeling...
Nunes-Correia, Isabel; Ramalho-Santos, João; Nir, Shlomo; Lima, Maria C. Pedroso deWe performed a detailed kinetic analysis of the uptake of influenza virus (A/PR8/34) by Madin Darby canine kidney (MDCK) cells in culture. Experimental procedures were based on the relief of fluorescence self-quenching of the fluorescent probe octadecylrhodamine B chloride (R18) incorporated in the viral envelope. Equilibrium for binding of influenza virus to MDCK cells (2.5 × 106/mL) was reached quicker with t...
Sendai Virus Fusion Activity as Modulated by Target Membrane Components
Nunes-Correia, Isabel; Ramalho-Santos, João; Pedroso de Lima, Maria C.We have studied the differences between erythrocytes and erythrocyte ghosts as target membranes for the study of Sendai virus fusion activity. Fusion was monitored continuously by fluorescence dequenching of R18-labeled virus. Experiments were carried out either with or without virus/target membrane prebinding. When Sendai virus was added directly to a erythrocyte/erythrocyte ghost suspension, fusion was always...
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