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Transformation of rodent fibroblasts by herpes simplex virus: Presence of morphological transforming region 1 (MTR 1) is not required for the maintenance of the transformed state
Author(s) -
Bauer Georg,
Kahl Susanne,
Sawhney Iva Singh,
Höfler Petra,
Gerspach Ralph,
Matz Bertfried
Publication year - 1992
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910510515
Subject(s) - biology , herpes simplex virus , transformation (genetics) , dna , virus , gene , viral transformation , viral replication , mutant , dna replication , virology , microbiology and biotechnology , cell culture , genetics
Studies on the mechanisms of transformation of mammalian cells by herpes simplex virus (HSV) in vitro have been prevented so far by the extremely low transformation frequencies obtained in monolayer culture. Here we present a transformation system that relies on the direct seeding in soft agar of infected single cells, thus avoiding negative interactions between normal and transformed cells. We took advantage of HSV‐1 temperature‐sensitive mutants at the UL9 locus, which codes for a DNA‐binding protein necessary for viral DNA replication. At the non‐permissive temperature, viral DNA synthesis and late gene expression are prevented. Viral gene expression is restricted to immediate early and early genes. Induction of transformation was highly efficient in our one‐step transformation system. It depended on intact viral particles and viral DNA. Immediate early and/or early viral gene expression was sufficient to induce transformation. Colonies were stably transformed and did not show any rescue of viable virus after temperature downshift and co‐cultivation with susceptible cells. Transformed cells maintained the transformed state in the absence of viral DNA. Our data therefore support the “hit‐and‐run” hypothesis for the transforming effect of HSV. © 1992 Wifey‐Liss, Inc.