Migration of vesicular stomatitis virus glycoprotein to the nucleus of infected cells.
A new means of direct visualization of the early events of viral infection by selective fluorescence labeling of viral proteins coupled with digital imaging microscopy is reported. The early phases of viral infection have great importance for understanding viral replication and pathogenesis. Vesicular stomatitis virus, the best-studied rhabdovirus, is composed of an RNA genome of negative sense, five viral proteins, and membrane lipids derived from the host cell. The glycoprotein of vesicular stomatitis virus was labeled with fluorescein isothiocyanate, and the labeled virus was incubated with baby hamster kidney cells. After initiation of infection, the fluorescence of the labeled glycoprotein was first seen inside the cells in endocytic vesicles. The fluorescence progressively migrated to the nucleus of infected cells. After 1 h of infection, the virus glycoprotein was concentrated in the nucleus and could be recovered intact in a preparation of purified nuclei. These results suggest that uncoating of the viral RNA occurs close to the nuclear membrane, which would precede transcription of the leader RNA that enters the nucleus to shut off cellular RNA synthesis and DNA replication.
Da Poian
A T
AT
Departamento de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Brazil.
Gomes
A M
AM
Oliveira
R J
RJ
Silva
J L
JL
D000818:Animals; D002451:Cell Compartmentation; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D006224:Cricetinae; D004705:Endocytosis; D006023:Glycoproteins / Q000378:metabolism; D006874:Hydrostatic Pressure; D008562:Membrane Glycoproteins; D008856:Microscopy, Fluorescence; D011487:Protein Conformation; D013997:Time Factors; D014721:Vesicular stomatitis Indiana virus / Q000254:growth & development*; D014759:Viral Envelope Proteins / Q000378:metabolism*; D014779:Virus Replication
C028391:G protein, vesicular stomatitis virus; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins