Detection of a protein encoded by the vaccinia virus C7L open reading frame and study of its effect on virus multiplication in different cell lines

• Published in: Journal of general virology 1993-07, Vol.74 (7), p.1409-1413
• Main Authors: Oguiura, Nancy, Spehner, Daniele, Drillien, Robert
• Format: Article
• Language: English
• Subjects: Animals; beta-Galactosidase - biosynthesis; beta-Galactosidase - genetics; Biological and medical sciences; Cell Line; Cloning, Molecular; Cricetinae; DNA Replication; Fundamental and applied biological sciences. Psychology; Gene Deletion; Genes, Bacterial; Genome, Viral; Humans; Microbiology; Miscellaneous; Open Reading Frames; Phenotype; Recombinant Fusion Proteins - biosynthesis; Recombination, Genetic; Restriction Mapping; Tumor Cells, Cultured; Vaccinia virus - genetics; Vaccinia virus - physiology; Viral Proteins - biosynthesis; Virology; Virus Replication
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-74-7-1409

Unité INSERM 74 and Laboratoire Commun ULP-Synthélabo, Institut de Virologie de la Faculté de Médecine, 3 rue Koeberlé, 67000 Strasbourg, France Vaccinia virus encodes several proteins, the activity of which is essential for multiplication in different cell types. Both the C7L and K1L open reading frames (ORFs) have been characterized as viral determinants for multiplication in human cells. To confirm and extend these findings we inserted the C7L ORF into the genome of a mutant virus unable to multiply in human cells and showed that this virus recovered its ability to replicate. Deletion of C7L from a wild-type viral genome did not adversely affect virus multiplication in human cells but it did reduce replication in hamster Dede cells. When both C7L and K1L were deleted from the vaccinia virus genome only poor or no viral yields were obtained from various human cell lines. Recombinant viruses were also constructed to facilitate the study of C7L protein synthesis during infection. One virus in which the lacZ ORF was fused downstream and in-frame with the C7L ORF enabled us to characterize the C7L protein as an early gene product. Another recombinant virus was constructed so that the carboxy terminus of the C7L ORF product contained an additional 28 amino acids from the carboxy terminus of K1L. Tagging of C7L in this way allowed us to detect the fusion protein by immunoprecipitation with antibodies against the K1L protein. Furthermore, the hybird protein retained its biological properties. The recombinant viruses constructed in this work should be useful for studies of the molecular basis of the activity of viral host range proteins. Present address: Laboratorio de Genética, Instituto Butantan, Av. Vital Brazil 1500, 05504 Sao Paulo, SP Brazil. Received 15 December 1992; accepted 5 February 1993.