Patterns of transient expression of the arenavirus nucleocapsid protein gene in transfected cells.
The cloned genes for the nucleocapsid proteins N of Junín and LCM (lymphocytic choriomeningitis) arenaviruses were inserted into the SV40-derived expression vector designated pKG4. When BHK-21 (baby hamster kidney fibroblasts) and CV-1 (African green monkey kidney fibroblasts) cell lines were transfected using these constructions, the transient expression yielded a polypeptide that could not be distinguished either by size nor by immunoreactivity from the N protein synthesized during the viral infection. The immunofluorescence analysis showed a pattern of intracellular localization similar to that observed in virus infected cells, i.e. varying from a diffuse cytoplasmic staining to granules, either distributed throughout the cytoplasm or concentrated in the perinuclear region. The association of the N protein with basophilic granules is similar to that observed in the cytopathic effect caused by arenaviruses, and could be related to the physicochemical properties of this polypeptide containing numerous basic amino acid sequences, that would allow for the interaction with cellular RNAs.
Rivera Pomar
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RV
Area de Química Biológica y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Argentina.
Manzella
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EM
Ghiringhelli
P D
PD
Grau
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O
Romanowski
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V
D000818:Animals; D006477:Arenaviruses, New World / Q000235:genetics*; D002213:Capsid / Q000096:biosynthesis*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003588:Cytopathogenic Effect, Viral; D003594:Cytoplasmic Granules / Q000378:metabolism / Q000648:ultrastructure; D005347:Fibroblasts / Q000378:metabolism / Q000648:ultrastructure; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D008217:Lymphocytic choriomeningitis virus / Q000235:genetics*; D008647:Mesocricetus; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D013539:Simian virus 40; D014162:Transfection; D014758:Viral Core Proteins / Q000096:biosynthesis*
D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins