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Biology of vaccinia virus acylproteins

Dennis E Hruby
1998 Frontiers in Bioscience  
Additionally, Child and Hruby have demonstrated the existence of at least five more palmitylproteins induced by VV (27) .  ...  The identification of the 92-kDa palmitylprotein as the ATI protein was made by Martin and Hruby as discussed above, but the 14-and 17-kDa palmitylproteins have not yet been identified.  ... 
doi:10.2741/a280 fatcat:pnc3wdvkrjexlgg7uvwpgmm3tu

Identification and Analysis of Vaccinia Virus Palmitylproteins

Douglas W. Grosenbach, Scott G. Hansen, Dennis E. Hruby
2000 Virology  
E-mail:  ...  At 4 hpi (also 4 h posttransfection) an additional 500 l of MEM-E, LG/GS was added. If label was to be added, it was added with the 500 l of MEM-E, LG/GS at 4 hpi.  ... 
doi:10.1006/viro.2000.0522 pmid:11017799 fatcat:lg32dpue7rb2tffc6fh7dakjsy

Importance of disulphide bonds for vaccinia virus L1R protein function

Robert E Blouch, Chelsea M Byrd, Dennis E Hruby
2005 Virology Journal  
G4L is likened to the downstream thioredoxin-like proteins DsbA in E. coli, and PDI and its homologues in the ER of yeast.  ...  Transfections of plasmid DNA were performed using 2 µg of pUC19, p(E/ Lp)L1R or p(wtp)L1R. All infections were harvested at 24 hpi and titered on BSC 40 cells.  ... 
doi:10.1186/1743-422x-2-91 pmid:16336686 pmcid:PMC1318495 fatcat:usbo3yhl5jgphdiwt5ecamouga

Development of ST-246® for Treatment of Poxvirus Infections

Robert Jordan, Janet M. Leeds, Shanthakumar Tyavanagimatt, Dennis E. Hruby
2010 Viruses  
ST-246 (Tecovirimat) is a small synthetic antiviral compound being developed to treat pathogenic orthopoxvirus infections of humans. The compound was discovered as part of a high throughput screen designed to identify inhibitors of vaccinia virus-induced cytopathic effects. The antiviral activity is specific for orthopoxviruses and the compound does not inhibit the replication of other RNA-and DNA-containing viruses or inhibit cell proliferation at concentrations of compound that are antiviral.
more » ... ST-246 targets vaccinia virus p37, a viral protein required for envelopment and secretion of extracellular forms of virus. The compound is orally bioavailable and protects multiple animal species from lethal orthopoxvirus challenge. Preclinical safety pharmacology studies in mice and non-human primates indicate that ST-246 is readily absorbed by the oral route and well tolerated with the no observable adverse effect level (NOAEL) in mice measured at 2000 mg/kg and the no observable effect level (NOEL) in non-human primates measured at 300 mg/kg. Drug substance and drug product processes have been developed and commercial scale batches have been produced using Good Manufacturing Processes (GMP). Human phase I clinical trials have shown that ST-246 is safe and well tolerated in healthy human volunteers. Based on the results of the clinical evaluation, once a day dosing should provide plasma drug exposure in the range predicted to be antiviral based on data from efficacy studies in animal models of orthopoxvirus disease. These data support the use of ST-246 as a therapeutic to treat pathogenic orthopoxvirus infections of humans.
doi:10.3390/v2112409 pmid:21994624 pmcid:PMC3185582 fatcat:fkyj5ttwr5cupkiuez2xc7g2qa

A host cell membrane protein, golgin-97, is essential for poxvirus morphogenesis

Dina Alzhanova, Dennis E. Hruby
2007 Virology  
In contrast, depletion of golgin-97 resulted in the loss of MV particles and dramatic increase in the number of IVs (Figs. 3B, C, E-I).  ...  HeLa cells transfected with negative RNAi control (A and D), dsRNA-1 (B, E, G-I) or dsRNA-2 (C and F) were infected with VV WR at an MOI of 5.0 pfu/cell.  ... 
doi:10.1016/j.virol.2007.01.003 pmid:17276477 pmcid:PMC2680701 fatcat:z22b4h2po5gidgdvkkom2wwzaq

Vaccinia Virus Replication I. Requirement for the Host-Cell Nucleus

Dennis E. Hruby, Linda A. Guarino, Joseph R. Kates
1979 Journal of Virology  
(E) Mature virus particle, 12 h. Bar = 0.1 ALM. FIG. 6 . 6 Electron micrographs of thin sections of vaccinia virus-infected BSC-40 cytoplasts.  ... 
doi:10.1128/jvi.29.2.705-715.1979 fatcat:ffga42w7zbhdrm7sfwqjr7nud4

A tandemly–oriented late gene cluster within the vaccinia virus genome

Scott L. Weinrich, Dennis E. Hruby
1986 Nucleic Acids Research  
0 1 G L J H D A B 20kbp SalI SalI BamHi DIA 11 BMI E SR Eo EcoRIIjEcoRI E.oRl idIIIs a Sail BmnHI EcoRI EcoR Hi,dIN Ss BIkbp 5115bp 1647 bp 0 Figure 1.  ...  major structural polypeptides just upstream of this region (18, 29) , the observation that several different VV temperature-sensitive mutants are marker rescued by restriction fragments of this region (E.  ... 
doi:10.1093/nar/14.7.3003 pmid:3008103 pmcid:PMC339717 fatcat:cy3wxfkcj5ezfizna6sswginqu

Chlamydial development is blocked in host cells transfected with Chlamydophila caviae incA

Damir Alzhanov, Jennifer Barnes, Dennis E Hruby, Daniel D Rockey
2004 BMC Microbiology  
Jennifer Barnes and Dennis Hruby designed and performed the work with the Vaccinia virus recombinants.  ...  Panel E; HeLa cell transfected with pCtAWT and subsequently infected at an MOI of 0.25 with C. caviae.  ... 
doi:10.1186/1471-2180-4-24 pmid:15230981 pmcid:PMC459217 fatcat:yydlxclcnjaavonicnou6kxbcm

Mutational analysis of the potential catalytic residues of the VV G1L metalloproteinase

Kady M Honeychurch, Chelsea M Byrd, Dennis E Hruby
2006 Virology Journal  
The essential downstream E residue for M16 MMPs is found within a region containing several completely conserved E residues [23] .  ...  A B E110 is the downstream E residue involved in zinc coordination (Fig. 2 ).  ... 
doi:10.1186/1743-422x-3-7 pmid:16504157 pmcid:PMC1420270 fatcat:ojkn2qahmnbktgxy2byq6mxzxm

Palmitylation of the Vaccinia Virus 37-kDa Major Envelope Antigen

Douglas W. Grosenbach, David O. Ulaeto, Dennis E. Hruby
1997 Journal of Biological Chemistry  
At 6 HPI, the culture supernatant was aspirated and the cells washed with warm MEM-E.  ...  The wash was aspirated and replaced with 1 ml of MEM-E containing 200 Ci of [9,10-3 H]palmitic acid ([ 3 H]PA, DuPont NEN) or 1 ml of MEM-E containing 200 Ci of [9,10-3 H]oleic acid ([ 3 H]OA, DuPont NEN  ...  Ulaeto and Dennis E.  ... 
doi:10.1074/jbc.272.3.1956 pmid:8999886 fatcat:qks2cchzenhihalg4557lucdty

Development of the small-molecule antiviral ST-246®as a smallpox therapeutic

Douglas W Grosenbach, Robert Jordan, Dennis E Hruby
2011 Future Virology  
In each of the E-IND cases described here, disease occurred as a result of smallpox vaccination in a setting of immune dysregulation.  ...  A second E-IND was issued for a recent vaccinee who had received the smallpox vaccine in preparation for a military deployment [62] .  ... 
doi:10.2217/fvl.11.27 pmid:21837250 pmcid:PMC3151656 fatcat:ah22dpco3nbq7la2b5tiuti4tu

Discovery and development of antiviral drugs for biodefense: Experience of a small biotechnology company

Tove C. Bolken, Dennis E. Hruby
2008 Antiviral Research  
The unmet need for effective antivirals against potential agents of bioterrorism and emerging infections is obvious; however, the challenges to developing such drugs are daunting. Even with the passage of Project BioShield and more recently the BARDA legislation, there is still not a clear market for these types of drugs and limited federal funding available to support expensive drug development studies. SIGA Technologies, Inc. is a small biotech company committed to developing novel products
more » ... r the prevention and treatment of severe infectious diseases, with an emphasis on products for diseases that could result from bioterrorism. Through trials and error SIGA has developed an approach to this problem in order to establish the infrastructure necessary to successfully advance new antiviral drugs from the discovery stage on through to licensure. The approach that we have taken to drug development is biology driven and dependent on a dispersive development model utilizing essential collaborations with academic, federal, and private sector partners. This consortium approach requires success in acquiring grants and contracts as well as iterative communication with the government and regulatory agencies. However, it can work as evidenced by the rapid progress of our lead antiviral against smallpox, ST-246, and should serve as the template for development of new antivirals against important biological pathogens.
doi:10.1016/j.antiviral.2007.07.003 pmid:17765333 pmcid:PMC2972676 fatcat:2bs6xfpsjfag7p3znf6wnbgjb4

Analysis of vaccinia virus temperature-sensitive I7L mutants reveals two potential functional domains

Megan J Moerdyk, Chelsea M Byrd, Dennis E Hruby
2006 Virology Journal  
Dts-4 (D), Cts-16 (E), Cts-34 (F), Dts-8 (G), Dts-35 (H) and Dts-93 (I) at the non-permissive temperature of 41C. Bars represent 400 nm except in C, D and F (bar = 200 nm).  ... 
doi:10.1186/1743-422x-3-64 pmid:16945137 pmcid:PMC1570340 fatcat:xal2r5kxkngcrlyyq3digkbstm

Development of an in vitro cleavage assay system to examine vaccinia virus I7L cysteine proteinase activity

Chelsea M Byrd, Dennis E Hruby
2005 Virology Journal  
selected inhibitors of metalloproteinases, aspartic acid proteinases or serine proteinases (EDTA, pepstatin, and PMSF, respectively), but was sensitive to several general cysteine proteinase inhibitors (E-  ...  N o E-64 100 µM Y e s EST 10 µM N o EST 100 µM Y e s IA 1 mM Yes Leupeptin 1 mM No NEM 2.5 mM Yes TTP inhibitors TTP-6171 50 µM Y e s TTP-6171 20 µM Y e s TTP-1021 50 µM Y  ...  e s TTP-1021 20 µM Y e s TTP-0961 50 µM N o TTP-0961 20 µM N o (page number not for citation purposes) Publish with Bio Med Central and every scientist can read your work free of  ... 
doi:10.1186/1743-422x-2-63 pmid:16105175 pmcid:PMC1198262 fatcat:swt73mlibrbepffaapkaee27ci

BiZyme: A Novel Fusion Protein-Mediating Selection of Vaccinia Virus Recombinants by Fluorescence and Antibiotic Resistance

Scott G. Hansen, Torrey A. Cope, Dennis E. Hruby
2002 BioTechniques  
As positive controls, E. coli CAT displayed 28.6% conversion, while pGNR.CAT-transfected cells had 94.8% conversion.  ...  CAT, a common antibiotic resistance gene found in bacterial species such as E. coli , was chosen for its readily detectable and assayable activity.  ... 
doi:10.2144/02325dd08 pmid:12019792 fatcat:zsfnajoj2zdc3fgoqalht4xiiu
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