On the cellular localization of the components of the herpes simplex virus type 1 helicase-primase complex and the viral origin-binding protein.
We constructed recombinant viruses based on the herpes simplex virus type 1 mutant tsK which individually were able to express the products of four viral DNA replication genes (UL5, UL8, UL9 and UL52) in the absence of any of the other proteins required for viral DNA synthesis. These viruses were used in immunofluorescence experiments to investigate the cellular localization of the four replication proteins expressed. The results demonstrated that all three components of the viral helicase-primase complex (UL5, UL8 and UL52 proteins) must be co-expressed to allow their efficient localization to the nucleus. Since the UL5 and UL52 proteins together form a complex which is enzymatically indistinguishable from a complex formed from all three proteins, a possible role of the UL8 protein may be in facilitating nuclear uptake. The UL9 protein (origin-binding protein) efficiently entered the cell nucleus when expressed alone. Both UL9 protein and the tripartite helicase-primase complex exhibited patterns of fluorescence which resembled the 'pre-replicative sites' described previously.
Calder
J M
JM
Medical Research Council Virology Unit, Institute of Virology, Glasgow, U.K.
Stow
E C
EC
Stow
N D
ND
D002467:Cell Nucleus / Q000382:microbiology; D004265:DNA Helicases / Q000235:genetics / Q000302:isolation & purification*; D019915:DNA Primase; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D014176:Protein Biosynthesis; D012316:RNA Nucleotidyltransferases / Q000235:genetics / Q000302:isolation & purification*; D011995:Recombination, Genetic; D018139:Simplexvirus / Q000737:chemistry*; D014764:Viral Proteins / Q000235:genetics / Q000302:isolation & purification*; D014779:Virus Replication
D004268:DNA-Binding Proteins; D014764:Viral Proteins; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; C062021:helicase-primase, Human herpesvirus 1; D004265:DNA Helicases