Comparison of JC and BK Human Papovaviruses with Simian Virus 40: Restriction Endonuclease Digestion and Gel Electrophoresis of Resultant Fragments

Abstract

JC virus was found to have a buoyant density of 1.20 g/cm3 in linear sucrose-D2O and 1.35 g/cm3 in cesium chloride isopycnic gradients. DNA extracted either from JC-infected cultures or from gradient-purified virions occupied a dense position relative to linear DNA in cesium chloride/ethidium bromide gradients, and the circular configuration of the extracted DNA was confirmed by electron microscopy, with a measured molecular weight of 2.93 x 106. DNA from BK virus was similarly prepared and compared to JC and to an SV40 DNA standard by digestion with restriction endonuclease preparations from Haemophilus influenzae, Haemophilus parainfluenzae, and Escherichia coli. Digests were electrophoretically analyzed on gradient polyacrylamide slab gels or agarose gels, and the three viruses were found to have distinctly different cleavage patterns by this form of analysis: JC and BK viruses were almost entirely different from SV40 and significantly different from each other. Thus, JC and BK human papovaviruses appear to be discrete new members of the papovavirus group, rather than SV40 variants.

Viral variant nucleotide sequences help expose leukocytic positioning in the JC virus pathway to the CNS

“…In 1974, restriction endonuclease comparisons of JCV, BKV, and SV40 DNA showed widely different digestion patterns, confirming evidence that these three polyomaviruses were truly discrete [24]. Polyomaviruses were, therefore, distinguishable by nucleic acid cohybridization techniques such as Southern blotting [25][26][27] and in situ hybridization [28,29].…”

Serological typing scheme for bk‐like isolates of human polyomavirus

“…It is well known that tissue-culture-grown stocks of BK contain DNA heterogeneous in size with many defective genomes [Osborn et al, 1974;Howley et al, 1975;Seehafer et al, 19751. In some isolates the fulllength genome may predominate, whereas in others, such as MG and IV in the present study, no distinct a x lo6 daltons.…”

Analysis of JC virus DNA purified directly from human progressive multifocal leukoencephalopathy brains

“…The yet we were able to purify 20 and 60 jig of viral NA strains DNA directly from 10-g samples of brain tissue t B (0.65 to from each case. This suggests that JCV is diffigment mea-cult to purify from the lipid-rich fractions of the JCV (Mad-brain or that there may be abundant nonencap-,er than the sidated viral DNA present in the infected brain 2 marked heterogeneity of JCV DNA species extracted from propagated virus or directly from infected cells in permissive cell systems (9,17,22,23,25,39). The degree of size heterogeneity among JCV DNA species seemed to depend upon the multiplicity of infection (22) and is thought to be due to the generation of defective, shortened DNA species during passage in tissue culture.…”

“…Recently, JCV was adapted to adult human brain cell culture (42). When JCV is grown in PHFG cells, the viral DNA may be heterogeneous in size, depending on such factors as the particular batch of cells or the multiplicity of infection of virus (9,17,22,25). Viral size has also been observed in human amnion cells infected with JCV (39) or in infected HEK cells (23).…”