A noninverting genome of a viable herpes simplex virus 1: presence of head-to-tail linkages in packaged genomes and requirements for circularization after infection.
The wild-type herpes simplex virus 1 genome consists of two components, L and S, which invert relative to each other, giving rise to four isomers. Previously we reported the construction of a herpes simplex virus 1 genome, HSV-1(F)I358, from which 15 kilobase pairs of DNA spanning the junction between L and S components were deleted and which no longer inverted (Poffenberger et al., Proc. Natl. Acad. Sci. U.S.A. 80:2690-2694, 1983). Further studies on the structure of HSV-1(F)I358 revealed the presence of two submolar populations among packaged DNA. The first, comprising no more than 10% of total packaged DNA, consisted of defective genomes with a subunit size of 36 kilobase pairs. The results suggest that this population arose by recombination through a directly repeated sequence inserted in place of the deleted L-S junction. The second minor population consisted of HSV-1(F)I358 DNA linked head-to-tail. Analyses of the structure of HSV-1(F)I358 DNA after infection indicated that the fraction of total DNA linked head-to-tail increased to approximately 40 to 50% within 30 min after exposure of cells to virus. The formation of head-to-tail linkages did not require de novo protein synthesis. Our interpretation of the results is that the termini of full-length DNA molecules are held together during packaging, that a small fraction of the termini is covalently linked during or after packaging, and that the remainder is covalently joined after the release of viral DNA from the infecting virus by either host or viral factors introduced into the cell during infection.
Poffenberger
K L
KL
Roizman
B
B
D002460:Cell Line; D002467:Cell Nucleus / Q000382:microbiology; D003513:Cycloheximide / Q000494:pharmacology; D004270:DNA, Circular / Q000378:metabolism*; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000378:metabolism*; D005814:Genes, Viral; D006801:Humans; D010746:Phosphonoacetic Acid / Q000494:pharmacology; D014176:Protein Biosynthesis; D012091:Repetitive Sequences, Nucleic Acid; D018139:Simplexvirus / Q000235:genetics* / Q000378:metabolism / Q000502:physiology
D004270:DNA, Circular; D004274:DNA, Recombinant; D004279:DNA, Viral; D003513:Cycloheximide; D010746:Phosphonoacetic Acid