The incorporation of radioactive phosphorus in the influenza virus and its distribution in serologically active virus fractions

Abstract

1. When the D.S.P. strain of influenza virus A is grown in eggs into which 100 μc. of radioactive inorganic phosphate has been introduced the virus incorporates32P into its structure.2. Some 20–25% of the virus32P is found in the virus phospholipid; the remainder is combined with the virus protein and is probably present in the virus nucleic acid.3. When the virus is disintegrated by ether treatment with the liberation of separate red-cell agglutinating and complement-fixing ‘soluble antigen’ particles the non-lipid32P is found to be associated with the soluble antigen fraction and not with the haemagglutinin.4. It is suggested that the complement-fixing soluble antigen is a nucleoprotein while the haemagglutinin is a phosphorus-free protein.

The nucleic acid and carbohydrate content of influenza virus A and of virus fractions produced by ether disintegration

“…The complement-fixing unit was apparently identical with the soluble antigen found in infected tissues, and, by the use of virus labelled with radiophosphorus, Hoyle, Jolles & Mitchell (1954) showed that it carried the whole of the virus nucleic acid. The haemagglutinating particle was apparently a nucleic acid-free enzyme.…”

“…The whole of the nucleic acid of the infective virus particle appears to be carried by the soluble antigen (Hoyle, Jolles & Mitchell, 1954). Since the infective virus contains 0-74 % RNA the soluble antigen accounts for about 14 % of the weight of the infective particle, and a spherical infective particle of diameter 100m,t and a particle weight of 3 x 108 would contain about 70 molecules of soluble antigen.…”

The Disintegration of Influenza Virus Particles on Entry into the Host Cell. Studies with Virus Labelled with Radiophosphorus

“…By increasing the dose of 32p Hoyle et al (1954) and Liu et al (1954) produced virus preparations much more heavily labelled than those of Graham. For the purpose of the present work it was necessary to obtain very heavily labelled virus, and it was found that the dose of 32p could be increased to 1 millicurie per egg, and when this was done preparations of virus could be obtained in which the Geiger count per minute per ml.…”

“…The demonstration by Graham & McLelland (1949, Hoyle, Jolles & Mitchell (1954) and Liu, Blank, Spizizen & Henle (1954) that influenza virus could be labelled with radiophosphorus afforded a possible method of detecting the fate of the infecting particle on entry into the host cell. This paper describes the behaviour of 32p labelled virus introduced as a primary inoculum in fertile eggs and affords direct evidence that the infecting particle becomes disintegrated on entry into the host cell.…”

“…Most of the work in this paper has been done with virus labelled by the technique used by Hoyle et al (1954) with the modification that the dose of 32p per egg was increased from 100 microcurie to 1 millicurie. The technique consists of growing the virus in eggs inoculated into the allantoic sac with 1 millicurie of32p in the form of disodium phosphate, collecting the infected allantoic fluid and concentrating and purifying the virus by two cycles of adsorption-elution from guinea-pig red cells, the cells carrying the adsorbed virus being washed several times with ice-cold saline before elution to remove contaminating inorganic 32p.…”