UV irradiation analysis of complementation between, and replication of, RNA-negative temperature-sensitive mutants of Newcastle disease virus

Abstract

Random UV irradiation-induced lesions destroy the infectivity of Newcastle disease virus (NDV) by blocking downstream transcription from the single viral promoter. The nucleocapsid-associated polypeptides most likely to be involved in RNA synthesis are located at the extreme ends of the genome: NP and P are promoter proximal genes, and L is the most distal gene. We attempted to order the two temperature-sensitive (ts) RNA-negative (RNA-) mutant groups of NDV by determining the UV target sizes for the complementing abilities of mutants Al and El. After UV irradiation, El was unable to complement Al, a result compatible with the A mutation lying in the L gene. In contrast, after UV irradiation, Al was able to complement El for both virus production and viral protein synthesis, with a target size most consistent with the E mutation lying in the P gene. UVirradiated virus was unable to replicate as indicated by its absence in the yields of multiply infected cells, either as infectious virus or as particles with complementing activity. After irradiation, ts mutant BlAP, with a non-ts mutation affecting the electrophoretic mobility of the P protein, complemented El in a manner similar to Al, but it did not amplify the expression of AP in infected cells. This too is consistent with irradiated virus being unable to replicate despite the presence of the components needed for replication of El. At high UV doses, Al was able to complement El in a different, UV-resistant manner, probably by direct donation of input polypeptides. Multiplicity reactivation has previously been observed at high-multiplicity infection by UV-irradiated paramyxoviruses. In this case, virions which are noninfectious because they lack a protein component may be activated by a protein from irradiated virions. Peeples, L. L. Rasenas, and M. A. Bratt, submitted for publication). As yet, no definite as-965 on July 7, 2020 by guest

Treatment of blood with a pathogen reduction technology using ultraviolet light and riboflavin inactivates Ebola virus in vitro

“…Our in vitro results demonstrate that the UV+RB treatment efficiently reduces EBOV titers to below limits of detection in both serum and whole blood. UV light has the capacity to induce changes in nucleic acids such as dimer formation or strand breaks, resulting in loss of viral infectivity, but viral reactivation due to a repeat exposure to UV light or other stimulus is recognized in DNA viruses and both positive‐ and negative‐sense SS RNA viruses . This viral reactivation in the absence of RB occurs through host cell repair mechanisms that can rescue the viral infectivity .…”

Intracellular processing of the Newcastle disease virus fusion glycoprotein

“…NDV, Australia-Victoria strain, was propagated and purified as described previously (10). Strain Australia-Victoria is a virulent NDV strain that undergoes a productive infection of CE cells to produce infectious virions (1,2,10,21,28; Kaplan and Bratt, Abstr. Annu.…”

Monoclonal antibodies to newcastle disease virus: delineation of four epitopes on the HN glycoprotein

“…Immunization. Virus used as immunogen was diluted to 50 ,ug/ml in Dulbecco phosphate-buffered saline (PBS) (7) and inactivated by a 15-min exposure to UV irradiation from a Sylvania G15T8 germicidal lamp at a distance of 78 cm (31). A BALB/c mouse was primed intraperitoneally with S ,ug of intact NDV AV-WT in complete Freund adjuvant and boosted 1 month later with the same immunogen, but in incomplete adjuvant.…”