scholarly journals Recombinant Myxoma Virus Expressing Walleye Dermal Sarcoma Virus orfC Is Attenuated in Rabbits

Viruses ◽  
2020 ◽  
Vol 12 (5) ◽  
pp. 517
Author(s):  
Laura V. Ashton ◽  
Sandra L. Quackenbush ◽  
Jake Castle ◽  
Garin Wilson ◽  
Jasmine McCoy ◽  
...  
Keyword(s):  
Oncolytic Virus ◽  
Clinical Signs ◽  
Myxoma Virus ◽  
Future Studies ◽  
Virus Dissemination ◽  
Walleye Dermal Sarcoma Virus ◽  
Cancer Models ◽  
New Zealand White Rabbits ◽  
Sarcoma Virus ◽  
Dermal Sarcoma

The poxvirus, myxoma virus (MYXV) has shown efficacy as an oncolytic virus (OV) in some cancer models. However, MYXV replication within murine cancer models and spontaneous canine sarcomas is short-lived. In mice, successful treatment of tumors requires frequent injections with MYXV. We hypothesize that treatment of cancer with a recombinant MYXV that promotes apoptosis could improve the efficacy of MYXV. The orfC gene of walleye dermal sarcoma virus (WDSV), which induces apoptosis, was recombined into the MYXV genome (MYXVorfC). A marked increase in apoptosis was observed in cells infected with MYXVorfC. To ensure that expression of WDSV orfC by MYXV does not potentiate the pathogenesis of MYXV, we evaluated the effects of MYXVorfC inoculation in the only known host of MYXV, New Zealand white rabbits. Virus dissemination in rabbit tissues was similar for MYXVorfC and MYXV. Virus titers recovered from tissues were lower in MYXVorfC-infected rabbits as compared to MYXV-infected rabbits. Importantly, rabbits infected with MYXVorfC had a delayed onset of clinical signs and a longer median survival time than rabbits infected with MYXV. This study indicates that MYXVorfC is attenuated and suggests that MYXVorfC will be safe to use as an OV therapy in future studies.

scholarly journals Walleye dermal sarcoma virus reverse transcriptase is temperature sensitive

2002 ◽  
Vol 83 (6) ◽  
pp. 1361-1365 ◽  
Author(s):  
Sharon K. Fodor ◽  
Volker M. Vogt
Keyword(s):  
Human Immunodeficiency Virus ◽  
Reverse Transcriptase ◽  
Temperature Sensitive ◽  
Leukaemia Virus ◽  
Virus Particles ◽  
Walleye Dermal Sarcoma Virus ◽  
Immunodeficiency Virus ◽  
Sarcoma Virus ◽  
Dermal Sarcoma

Walleye dermal sarcoma virus (WDSV) is a piscine retrovirus that replicates naturally in fish at temperatures near 4 °C. The reverse transcriptase (RT) protein from virus particles isolated from walleye tumours was purified and biochemically characterized. Like the RT of the distantly related murine leukaemia virus, WDSV RT sediments as a monomer in the absence of template. It exhibits a K m of 22 μM for TTP in an assay with poly(rA) as a template and oligo(dT) as a primer. The enzyme is rapidly inactivated at temperatures greater than 15 °C. The ratio of RT activity at 15 °C to that at 4 °C is similar for WDSV and recombinant human immunodeficiency virus type 1, suggesting that, at least with this template, the fish enzyme is not specially adapted to function more efficiently in the cold.

scholarly journals Walleye Dermal Sarcoma Virus: OrfA N-Terminal End Inhibits the Activity of a Reporter Gene Directed by Eukaryotic Promoters and Has a Negative Effect on the Growth of Fish and Mammalian Cells

1999 ◽  
Vol 73 (10) ◽  
pp. 8884-8889 ◽  
Author(s):  
Z. Zhang ◽  
D. Martineau
Keyword(s):  
Reporter Gene ◽  
Mammalian Cells ◽  
Full Length ◽  
Skin Tumor ◽  
Accessory Gene ◽  
Walleye Dermal Sarcoma Virus ◽  
Negative Effect ◽  
Sarcoma Virus ◽  
Eukaryotic Promoters ◽  
Dermal Sarcoma

ABSTRACT Walleye dermal sarcoma virus (WDSV) is a fish retrovirus causing a skin tumor termed walleye dermal sarcoma, which develops and regresses on a seasonal basis. The WDSV genome contains three short open reading frames designated orfA, orfB, andorfC in addition to the viral structural genes,gag, pol, and env. orfAand orfB transcripts are detected in tumors by reverse transcription-PCR. Recently, OrfA, whose amino acid sequence is similar to that of cyclins A and D, has been shown to complement a cyclin-deficient yeast strain. We report that expression of the accessory gene orfA inhibited nonspecifically the activity of a reporter gene directed by various eukaryotic promoters. In addition, stable transfection with the wild-type orfAgenerated substantially fewer G418-resistant colonies in both fish and mammalian cells than the parent vector. An orfA mutant expressing only the first N-terminal 49 residues of the full-length protein had the same negative effect on the activity of the reporter gene and on the number of stably transfected colonies as the full-length OrfA. Thus, OrfA inhibits cell growth and/or causes cell death, and the first 49 N-terminal residues of this protein are sufficient to cause these negative effects.

scholarly journals Identification and Characterization of cis-Acting Elements Residing in the Walleye Dermal Sarcoma Virus Promoter

2004 ◽  
Vol 78 (14) ◽  
pp. 7590-7601 ◽  
Author(s):  
Brett W. Hronek ◽  
Ashley Meagher ◽  
Joel Rovnak ◽  
Sandra L. Quackenbush
Keyword(s):  
Cell Line ◽  
Nuclear Extract ◽  
Dnase I ◽  
Negative Regulator ◽  
Luciferase Reporter ◽  
Walleye Dermal Sarcoma Virus ◽  
Nuclear Extracts ◽  
Sarcoma Virus ◽  
Footprint Analysis ◽  
Dermal Sarcoma

ABSTRACT Walleye dermal sarcoma virus (WDSV) is a complex retrovirus found associated with tumors that appear and regress on a seasonal basis. There are quantitative and qualitative differences in the amount of virus expression between developing and regressing tumors. To understand the role of host cell factors in WDSV expression, DNase I footprint analysis, electrophoretic mobility shift assays (EMSA), and reporter gene assays were employed. DNase I footprint analysis of the U3 region of the WDSV long terminal repeat with nuclear extract prepared from a walleye cell line revealed protection of an Oct1, AP1, Whn, and two E4BP4 sites. Additionally, three regions that contained no putative transcription factor binding sites were protected. EMSA confirmed the specific binding of the protected sites and revealed three additional sites, NF1, AP3, and LVa, not protected in DNase I footprint analysis. Site-directed mutagenesis of the individual sites, in the context of a luciferase reporter plasmid, revealed that the NF1, Oct1, AP1, E4BP4#2, AP3, and LVa sites contributed to transcription activation driven by the WDSV U3 region. Mutation of Novel#2 resulted in an increase in luciferase activity, suggesting the Novel#2 site may function to bind a negative regulator of transcription. Anti-Jun and anti-Fos antiserum specifically inhibited protein-DNA complex formation, indicating the presence of c-Jun and c-Fos in the walleye cell nuclear extracts and their participation in binding to the AP1 site. Interestingly, degenerative 15-bp repeats found in the U3 region are differentially protected in DNase I footprint analysis by the walleye cell line nuclear extract and regressing-tumor nuclear extract. EMSA utilizing the 15-bp repeat probe revealed that there are similarities of binding with W12 cell and developing-tumor nuclear extracts and that the binding differs from that observed with regressing-tumor nuclear extract.

scholarly journals Transcriptional Analysis of Walleye Dermal Sarcoma Virus (WDSV)

Virology ◽  
1997 ◽  
Vol 237 (1) ◽  
pp. 107-112 ◽  
Author(s):  
Sandra L. Quackenbush ◽  
Donald L. Holzschu ◽  
Paul R. Bowser ◽  
James W. Casey
Keyword(s):  
Transcriptional Analysis ◽  
Walleye Dermal Sarcoma Virus ◽  
Sarcoma Virus ◽  
Dermal Sarcoma

scholarly journals Walleye Dermal Sarcoma Virus: Molecular Biology and Oncogenesis

Viruses ◽  
2010 ◽  
Vol 2 (9) ◽  
pp. 1984-1999 ◽  
Author(s):  
Joel Rovnak ◽  
Sandra L. Quackenbush
Keyword(s):  
Molecular Biology ◽  
Walleye Dermal Sarcoma Virus ◽  
Sarcoma Virus ◽  
Dermal Sarcoma

scholarly journals Nucleotide sequence and protein analysis of a complex piscine retrovirus, walleye dermal sarcoma virus.

1995 ◽  
Vol 69 (9) ◽  
pp. 5320-5331 ◽  
Author(s):  
D L Holzschu ◽  
D Martineau ◽  
S K Fodor ◽  
V M Vogt ◽  
P R Bowser ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Protein Analysis ◽  
Walleye Dermal Sarcoma Virus ◽  
Sarcoma Virus ◽  
Dermal Sarcoma
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