Amino acid Requirement of Adenovirus Multiplication

Abstract

SUMMARYThe absence from the medium of any of the 13 amino acids essential for cell growth has an inhibiting effect on the multiplication of adenovirus 9-I5 and adenovirus I in HeLa cell cultures. The inhibition is accentuated by previous amino acid starvation of the cultures. Whereas with arginine deprivation, the arginine pool inside the cells is at a minimum within 3o rain, the cells are assumed to adapt slowly to the new metabolic state, which is characterized by an increased 'turnover' of protein synthesis. With arginine deficiency and in Hanks' BSS some synthesis of virus and capsid proteins takes place. Quantitative and possibly qualitative differences between the influence of the various deficient media were observed.The experiments rule out DNA synthesis as a primary cause of the amino acid deficiency effect. They lead to the hypothesis that arginine deficiency inhibits the formation of an essential protein which is synthesized very late in the infectious cycle under complete MEM.

293SF Metabolic Flux Analysis during Cell Growth and Infection with an Adenoviral Vector

“…However, DNA synthesis in Inf B is still active after the peak in viral titer. Similarly, Wigand and Kumel (1978) saw a maximum in DNA production at around 20 h postinfection, which then dropped drastically. Usually DNA is stable and a drop in content may indicate that the cell integrity is affected.…”

On the mechanism of arginine requirement for adenovirus synthesis

“…The reason why incomplete virions are not assembled subsequent to the release from the arginine block, may be due to the absence of a critical protein required for assembly which is only synthesized during the early portion of the late phase of infection. Indeed recent studies with actinomycin D suggested that such a protein might be translated from an mRNA transcribed before 16 hours postinfection (18).…”

“…Our results confirm the finding that none of the viral polypeptides which are demonstrable by current SI)S-polyacrylamide gel electrophoresis techniques are inhibited by arginine deprivation. Interestingly, viral DNA synthesized during a short period of arginine deficiency is incorporated into virions subsequent to reversal of the arginine block, while such viruses incorporated only newly synthesized proteins eventhough large pools of proteins synthesized during deficiency were available (6,11,18). That amino acid starvation is not the only cause for the non-utilization of 'old' pools of proteins for virus assembly was demonstrated by the non-utilization of such 'old' protein pools when a temperature-sensitive assembly mutant was shifted to the permissive temperature (8).…”

“…If such extensive substitution were indeed the case it is surprising that PVII is nevertheless processed. However it is possible that PVII, or other arginine containing proteins, are not substituted extensively because arginine is made available through the degradation of cellular proteins (3,18).…”

“…The growth of adenoviruses is dependent on arginine more than on any other single amino acid (12,13,18). Synthesis of herpes virus, SV40, polyoma and reovirus also appear to require arginine for production of infectious progeny (for refs.…”