Acquisition of an additional internal cleavage site differentially affects the ability of pseudorabies virus to multiply in different host cells.
The translocation of the 325 leftmost bp of the genome of pseudorabies virus (PrV) to the internal junction between the L and S components confers upon the virus a growth advantage relative to wild-type PrV in chicken embryo fibroblasts (CEFs) and chickens and a growth disadvantage in rabbit kidney (RK) cells and mice. To clarify the molecular basis for the species-specific growth characteristics of the translocation mutants, we have compared several parameters of the virus growth cycle in CEFs and RK cells infected with wild-type PrV and with translocation mutants. The salient findings are as follows. (i) The synthesis of early-late and late proteins is not as effective in CEFs as it is in RK cells, and these proteins, in particular, the major capsid proteins, accumulate less abundantly in CEFs than in RK cells. (ii) Cleavage of concatemeric DNA to genome-size molecules is also not as effective in CEFs as it is in RK cells. (iii) The internal junction present in translocation mutants is a functional cleavage site. (iv) In RK cells, translocation mutants are hypercleaved and a significant proportion of the total viral DNA is cleaved into subgenomic fragments. (v) In CEFs infected with translocation mutants, subgenomic fragments also accumulate but most of the viral DNA remains in concatemeric form. A model which postulates that the cell-specific growth advantage or disadvantage of the translocation mutants is related to the presence of a second cleavage site within their genomes and is affected by the efficiency of cleavage of concatemeric DNA in particular infected cell types is presented. The significance of these findings as they relate to the evolution of herpesviruses with class 2- and class 3-like genomes is discussed.
Rall
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Department of Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
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Sugg
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Veach
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Ben-Porat
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D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis / Q000302:isolation & purification; D005347:Fibroblasts; D016679:Genome, Viral; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000502:physiology*; D007668:Kidney; D007700:Kinetics; D011817:Rabbits; D015183:Restriction Mapping; D014178:Translocation, Genetic; D014764:Viral Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D014779:Virus Replication
D004279:DNA, Viral; D014764:Viral Proteins