This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Liu, G Q Articles by Ginsberg, H S Search for Related Content PubMed PubMed Citation Articles by Liu, G Q Articles by Ginsberg, H S

 Previous Article  |  Next Article 

J Virol. 1985 March; 53(3): 920-925

A thermolabile mutant of adenovirus 5 resulting from a substitution mutation in the protein VIII gene.

ABSTRACT

The mutant adenoviruses H5sub304 and H5RIr were isolated sequentially from adenovirus 5 wild type by selection for the loss of EcoRI restriction endonuclease sites by Jones and Shenk (Cell 13:181-188, 1978). sub304 lacks the site at 84.0 map units (m.u.), and RIr lacks both that and the site at 75.9 m.u. A set of derivatives of RIr that lack the site at 75.9 m.u. accumulated virus more slowly at 38.8 or 39.5 degrees C than those with the site present, as measured by low-multiplicity passage or single-step replication cycles, respectively. Since the EcoRI site at 75.9 m.u. is predicted to lie in the gene encoding the precursor to virion polypeptide VIII (pVIII), the failure to accumulate virus rapidly could lie either in some step in processing and assembly of virions or in an increased virion thermolability. The latter possibility was shown to be the case, as all strains mutated at the EcoRI 75.9 m.u. site were extremely thermolabile in vitro, even at 37 degrees C. CsCl equilibrium density centrifugation of heated crude stocks of RIr and sub304 demonstrated that loss of infectivity in RIr was accompanied by physical disruption of virions. Polyacrylamide gel electrophoresis of infected cell extracts or of purified virions showed that pVIII of RIr had an apparent molecular weight that was slightly greater than that of sub304, and mature RIr and sub304 virions displayed polypeptide VIIIs which appeared to be of identical molecular weights. Nucleotide sequence analysis of RIr demonstrated that it contained a 9-base-pair (bp) substitution for 6 bp found in sub304, leading to a loss of the EcoRI site and a predicted insertion of a single amino acid. Comparison of the sequence of sub304 with the published sequence of adenovirus 2 revealed two changes, a single transversion at bp 1,722 and a bp deletion at 1,749, leading to the loss of a TaqI site. The predicted reading frame change would lead to a stop codon at bp 1,885. This raises the question of whether adenovirus 2 and adenovirus 5 use the same reading fame for pVIII.

This article has been cited by other articles:

• Vellinga, J., Van der Heijdt, S., Hoeben, R. C. (2005). The adenovirus capsid: major progress in minor proteins. J. Gen. Virol. 86: 1581-1588 [Abstract] [Full Text]  
• Kovacs, G. M., Davison, A. J., Zakhartchouk, A. N., Harrach, B. (2004). Analysis of the first complete genome sequence of an Old World monkey adenovirus reveals a lineage distinct from the six human adenovirus species. J. Gen. Virol. 85: 2799-2807 [Abstract] [Full Text]