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PMID: 4132673 Published · ppublish English Journal Article

Effect of ultraviolet light on mengovirus: formation of uracil dimers, instability and degradation of capsid, and covalent linkage of protein to viral RNA.

Journal of virology ·Vol. 13 ·No. 3 ·1974-03-00 ·Pages 729-39

Miller RL, Plagemann PG

Abstract

UV irradiation of purified mengovirus resulted in a very rapid inactivation of the infectivity of the virions (D(37) [37% survival dose] = 700 ergs/mm(2)) which correlated in time with the formation of uracil dimers in the viral RNA. During the first 2 min of irradiation, an average of 1.7 uracil dimers were formed per PFU of virus inactivated. Hemagglutination activity of the virions began to decrease only after a lag period of about 5 min and at a much lower rate (D(37) = 84,000 ergs/mm(2)). This decrease coincided in time with the appearance of altered proteins in the capsid and a structural change in the capsid. Although 10- to 20-min irradiated virions appeared intact in the electron microscope and sedimented at 150S in sucrose density gradients, the RNA of the virions became accessible to RNase and extractable by low concentrations of sodium dodecyl sulfate, and the virions broke down upon equilibrium centrifugation in CsCl gradients. During longer periods of irradiation (30 to 60 min), a progressively greater proportion of the virions were converted to 14S protein particles and 80S ribonucleoprotein particles composed of intact viral RNA and about 30% of the capsid proteins, alpha, beta, and gamma. Empty capsids were not detectable at any time during 60 min of irradiation, by which time disruption of the virions was complete. Irradiation of complete virions also resulted in an increased sedimentation rate of the viral RNA and in the covalent linkage to the viral RNA of about 1% of the total capsid protein in the form of heterogeneous low-molecular-weight polypeptides. The two observations seem to be causally related, since irradiation of isolated viral RNA did not result in an increase in sedimentation rate of the RNA, even though uracil dimer formation in viral RNA occurred at about the same rate and to the same extent whether intact virions or viral RNA were irradiated.

MeSH Terms
Animals Carbon Radioisotopes Carcinoma, Hepatocellular Cell Line Centrifugation, Density Gradient Hemagglutination Tests Liver Neoplasms Mengovirus/growth & development,immunology,metabolism,radiation effects Microscopy, Electron Nucleoproteins/metabolism,radiation effects Phosphotungstic Acid Protein Binding/radiation effects RNA, Viral/metabolism,radiation effects Radiation Effects Rats Staining and Labeling Time Factors Tritium Ultraviolet Rays Uracil/metabolism Viral Proteins/metabolism,radiation effects
Chemicals
Carbon Radioisotopes Nucleoproteins RNA, Viral Viral Proteins Tritium Phosphotungstic Acid Uracil
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Miller R L
Plagemann P G
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31 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
Published
1974-03-00
Pages
729-39
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC355360
Subset
IM
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