Characterization of an early temperature-sensitive and cytocidal double mutant of adenovirus type 2

The Journal of General Virology
J C D'HalluinP Boulanger

Abstract

Human adenovirus type 2 mutant, H2 ts 111, presented a double phenotype: temperature-sensitive (ts) for initiation and elongation of DNA synthesis, and cytocidal (cyt) by its large-plaque formation and the nucleolytic cleavage of both viral and cellular DNAs. Both characters were recessive since they were efficiently complemented by wild-type or other DNA-negative ts mutants. H2 ts 111 DNA--terminal protein complex formed at 33 degrees C and chased at 39.5 degrees C showed a decreased affinity for glass fibre filters, concurrently with the loss of protein-linked DNA ends. H2 ts 111 DNA breakdown occurring upon shift-up to 39.5 degrees C therefore appeared to start in close proximity to the genome extremities. Marker rescue experiments showed that the ts character was abolished by co-infection with plasmid recombinants containing whole or part of the E2A region, encoding for the 72K DNA-binding protein. The N-terminal domain of this 72K protein has been assigned between 0 and 200 amino acids, and is supposed to have a function in late transcription control. Since H2 ts 111 mapped between 0 and 300 residues (63.6 to 65.9 map units), its mutation was most likely located between 200 and 300 amino acids, namely in the C-terminal dom...Continue Reading

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