"These findings indicate that the minor capsid proteins are packaged inside the capsid particle with viral genomic DNA. Our previous study demonstrated that the first 12 amino acids at the N-terminus of JCV VP1 are responsible for DNA binding and DNA packaging (Ou et al, 2001b). According to the simulated three-dimensional structure of JCV VP1 (Ou et al, 2001a), the N-terminus of the VP1 polypeptide is cryptic in the capsid. "
[Show abstract][Hide abstract] ABSTRACT: To investigate the DNA binding activity of the JC virus minor capsid protein, VP2, both wild-type and mutant VP2 were cloned and expressed in Escherichia coli. Southwestern blotting was employed for the DNA-binding assay. The results showed that VP2 was able to bind to DNA, except when either the last 13 or the last 29 amino acids were truncated. The results indicate that the DNA-binding domain of VP2 is located within the last 13 amino acids. Furthermore, we also demonstrated that Lys(332) and Lys(336) within the DNA-binding domain are crucial for DNA binding. The findings may provide further information for understanding the mechanism of virion assembly of the JC virus.
[Show abstract][Hide abstract] ABSTRACT: Human JC virus (JCV) belongs to the family of Polyomaviridae. The viral capsid is composed of 72 capsomeres. Five VP1 molecules make up a capsomere structure. To investigate the minimal sequences on JCV VP1 polypeptide required for capsid assembly, the first 12 (Delta N12) and 19 (Delta N19) amino acids at the N-terminus and the last 16 (Delta C16), 17 (Delta C17), and 31 (Delta C31) amino acids at the C-terminus of VP1 were truncated and expressed in E. coli. The VP1 proteins of Delta N12 and Delta C16 were able to self-assemble into a virus-like particle similar to that of wild-type (WT) VP1. However, the mutant proteins of Delta N19, Delta C17, and Delta C31 formed a pentameric capsomere structure as demonstrated by a 10-50% sucrose gradient centrifugation and electron microscopy. These results suggest that the 12 amino-terminal and 16 carboxy-terminal amino acids of VP1 are dispensable for the formation of virus-like particles, and further truncation at either end of VP1 leads to the loss of this property.
Journal of NeuroVirology 09/2001; 7(4):298-301. DOI:10.1080/13550280152537139 · 2.60 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Human JC virus (JCV) is a neurotropic virus, and the etiological agent of progressive multifocal leukoencephalopathy (PML), a fatal neurological disease. Because of its natural infection tropism, it is possible to use the JCV capsid as a gene-transducing vector for therapeutic purposes in neurological disorders. In the current study, a recombinant JCV virus-like particle (VLP) was generated and purified from yeast. VLP was able to accommodate and protect DNA molecules of up to approximately 2000 bp in length. VLP was able to package and deliver an antisense oligodeoxynucleotide (AS-ODN) against simian virus 40 (SV40) large tumor antigen (LT) into SV40-transformed human fetal glial (SVG) cells in order to inhibit expression of the oncoprotein. Subsequently, apoptosis of VLP-AS-ODN-treated cells was demonstrated after the blocking of LT expression. In addition, JCV VLP was able to deliver ODN into human astrocytoma, neuroblastoma, and glioblastoma cells with high efficiency. In vivo delivery of ODN into a human neuroblastoma tumor nodule by VLP was also demonstrated. These findings suggest that JCV VLP is a gene delivery vector with potential therapeutic use for human neurological disorders.
Human Gene Therapy 12/2004; 15(11):1077-90. DOI:10.1089/hum.2004.15.1077 · 3.76 Impact Factor
Data provided are for informational purposes only. Although carefully collected, accuracy cannot be guaranteed. The impact factor represents a rough estimation of the journal's impact factor and does not reflect the actual current impact factor. Publisher conditions are provided by RoMEO. Differing provisions from the publisher's actual policy or licence agreement may be applicable.