Herpes simplex virus DNA replication: the UL9 gene encodes an origin-binding protein.
Herpes simplex virus 1 contains seven genes that are necessary and sufficient for origin-dependent DNA synthesis in cultured cells. We have expressed the product of one of these genes, UL9, in insect cells by using a baculovirus expression vector. The apparent size of the UL9 protein, both in insect cells and in herpes simplex virus-infected Vero cells, is 82,000 Da. By using an immunoassay for protein-DNA interaction, we have shown that UL9 protein binds specifically to the herpes simplex virus origins of DNA replication, oriS and oriL. DNase I "footprint" analysis has shown that the UL9 protein interacts with two related sites on oriS, located on each arm of a nearly perfect palindrome. Our data strongly suggest that the origin-binding activity described previously by Elias et al. [Elias, P., O'Donnell, M. E., Mocarski, E. S. & Lehman, I. R. (1986) Proc. Natl. Acad. Sci. USA 83, 6322-6326] is the product of the UL9 gene.
Olivo
P D
PD
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Nelson
N J
NJ
Challberg
M D
MD
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I; D005786:Gene Expression Regulation; D005814:Genes, Viral; D005822:Genetic Vectors; D007118:Immunoassay; D007304:Insect Viruses / Q000235:genetics; D007313:Insecta; D008969:Molecular Sequence Data; D011485:Protein Binding; D018139:Simplexvirus / Q000235:genetics* / Q000502:physiology; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D014779:Virus Replication
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; D003850:Deoxyribonuclease I