Immunocytochemical localization of vesicular stomatitis virus proteins N and NS with monoclonal antibodies.
The purpose of this paper is to describe the immunocytochemical-localization of N and NS nucleocapsid proteins of vesicular stomatitis virus in the cells throughout the infectious cycle. N protein was detected in the cytoplasm at 2 h after infection and formed small cytoplasmic clusters which progressively increased in size and number. At 5-6 h, it formed large cytoplasmic inclusions. NS protein was detected in the cytoplasm a little later than N protein and showed almost the same immunostaining pattern. However, diffuse background staining of NS protein was identified throughout the cytoplasm by double immunostaining methods. At electron microscopic level, N protein was mostly granular and occasionally organized in strands at 2-3 h. At 5-6 h, numerous immunostained reaction products were organized in strands. The reaction products of NS protein were almost the same as those of N protein with the exception that diffuse background staining was observed. Cos cells, transfected with SV40 vector containing N gene obtained by recombinant DNA technique, showed clusters of N protein, but virtually no strand at electron microscopic levels. The rapid-freezing and deep-etching replica method demonstrated that loosely coiled VSV genome coated with N protein was localized on cytoplasmic sides of cell membranes in the infected cells. These results showed that complete virus genome replication was needed for strand formation of N and NS proteins and suggested that they were bound to VSV genomes in the infected cells.
Ohno
S
S
Arnheiter
H
H
Dubois-Dalcq
M
M
Lazzarini
R A
RA
D000818:Animals; D000911:Antibodies, Monoclonal; D002213:Capsid / Q000032:analysis; D002460:Cell Line; D002522:Chlorocebus aethiops; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D006651:Histocytochemistry; D007124:Immunoenzyme Techniques; D008854:Microscopy, Electron; D014721:Vesicular stomatitis Indiana virus / Q000032:analysis* / Q000235:genetics; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D017361:Viral Nonstructural Proteins; D014764:Viral Proteins