[PDF] Top 20 Mutational Analysis of the Herpes Simplex Virus Type 1 DNA Packaging Protein UL33

... herpesvirus DNA is cleaved and packaged but is then either lost from capsids or retained in capsids that do not protect the DNA from digestion with ...the packaging signal can be observed in samples ... See full document

... of protein synthesized by transrepression-competent mutants of ...of protein that would be detectable by Western blot analyses; alternatively, the expression of these mutants could be tightly controlled ... See full document

... all herpes- viruses. Herpes simplex virus (HSV) capsid pentons and hex- ons form spontaneously from five and six molecules of ICP5, respectively; these capsomeres are linked by triplexes ... See full document

... wt virus A, B, and C capsids. To determine whether UL17 was a capsid protein, BHK cells were infected either with a VP19C null mutant virus, which does not produce capsids, or with wt virus, ... See full document

... amplicon DNA rel- atively more efficiently than it packages its own genome, (iv) the two termini of packaged amplicons are present in similar amounts, and (v) neither packaged viral genomes nor ampli- cons are ... See full document

... for 1 h before being infected with 5 PFU/cell of v ⌬ ...blotting. Protein samples were separated on a 10% sodium dodecyl sulfate-polyacrylamide gel and detected by enhanced chemiluminescence using mAb02040 ... See full document

... h. Virus capsids were isolated and purified from nuclear lysates of infected cells on a 20 to 50% sucrose gradient, essentially as described previously ...7.4], 1 mM EDTA). Protein profiles were ... See full document

... of herpes simplex virus type I (HSV-1), consisting of UL5, UL8, and UL52, possesses 5 ⴕ to 3 ⴕ helicase, single-stranded DNA (ssDNA)-dependent ATPase, primase, and DNA ... See full document

... wild-type virus and a virus with a mutant version of cleav- age and packaging gene UL6, capsids localize exclusively to replication compartments, cells infected with the UL32 mutant exhibit an ... See full document

... for packaging of mature, linear ...stein-Barr virus (30). Alkaline nuclease has been found in a protein complex bound to the cleavage site of the HSV-1 ge- nome ...its DNA to nearly ... See full document

... The analysis of herpesvirus cleavage packaging signals has employed two principal ...of DNA replication and putative packag- ing signal (so-called amplicons) are transfected into tissue cul- ture ... See full document

... While it has been clear for a long time and has been reconfirmed in the accompanying report 11 that such assays of HSV promoter activity do not always accurately reflect kinetics or iden[r] ... See full document

... a DNA binding protein activates ...that DNA binding is essential for the activating functions of ICP4 (1, 44, ...However, mutational alteration of the binding sites in and around the ... See full document

... UL25 protein ranges from 80% to ...UL25 protein, we suspect that the single-triplex and UL25 binding site employed by the UL17 protein may result in greater loss during capsid purification, and ... See full document

... nucleotide 1 (14), and at the EcoRI site that occurs in the linker insertion in these ...dues 1 to 163, was joined to the C-terminal portion of mutant S13 from residues 264 to ...residues 1 to ...of ... See full document

... Cluster 1 is located in the apical region of the protein lo- cated on the same face as the view shown in ...cluster 1, traverses laterally across the entire molecule and abuts disordered loop ... See full document

... wild-type virus HSV-1(F) and the mutant HSV-7202 have been previously described (6, 16), and their titers were determined on Vero cell ...deletion virus HSV-gCB was determined by propagation ... See full document

... herpesvirus DNA. By introducing single copies of test packaging signals into an HSV-1 genome lacking the normal resident copies, we have overcome one of the major drawbacks of the previously used ... See full document

... A virus mutant was created by in vivo Red/ET recombination, using a Counter-Selection BAC modification kit sup- plied by Gene ...(Table 1) in a PCR with an rpsL-neo cassette as the DNA template, was ... See full document

... UL11 packaging. To compare the relative levels of packaging for wt and mutant derivatives of UL11, a transfection/infection-based assay was ...of DNA per 100-mm plate for each con- struct) using ... See full document