scholarly journals Transgenic Expression of Walleye Dermal Sarcoma Virus rv-cyclin (orfA) in Zebrafish does not Result in Tissue Proliferation

2010 ◽  
Vol 13 (2) ◽  
pp. 142-150 ◽  
Author(s):  
Thomas A. Paul ◽  
Joel Rovnak ◽  
Sandra L. Quackenbush ◽  
Kathleen Whitlock ◽  
Huiqing Zhan ◽  
...  
Keyword(s):  
Transgenic Expression ◽  
Tissue Proliferation ◽  
Walleye Dermal Sarcoma Virus ◽  
Sarcoma Virus ◽  
Dermal Sarcoma

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 PCR and RT-PCR Analysis of Infection and Transcriptional Activity of Walleye Dermal Sarcoma Virus (WDSV) in Organs of Adult Walleyes (Stizostedion vitreum)

1996 ◽  
Vol 33 (1) ◽  
pp. 66-73 ◽  
Author(s):  
F. M. Poulet ◽  
P. R. Bowser ◽  
J. W. Casey
Keyword(s):  
Viral Infection ◽  
Transcriptional Activity ◽  
Stizostedion Vitreum ◽  
Viral Dna ◽  
Tumor Bearing ◽  
Dna And Rna ◽  
Quantitative Estimates ◽  
Walleye Dermal Sarcoma Virus ◽  
Sarcoma Virus ◽  
Dermal Sarcoma

The pathogenesis of walleye dermal sarcoma virus (WDSV) infection was investigated in adult walleyes ( Stizostedion vitreum). Three tumor-bearing and three tumor-free walleyes were collected in the spring from Oneida Lake, New York, and analyzed for viral infection and transcriptional activity. Specifically, the target organs for viral infection and supporting viral transcriptional activity were determined by assessing for the presence of WDSV DNA and RNA in the brain, liver, kidney, skin, and spleen. For each organ, WDSV DNA and RNA were detected using the polymerase chain reaction (PCR) and reverse transcription PCR (RTPCR) respectively. Quantitative estimates of the number of viral DNA and RNA copies were obtained in each case by comparing the signal intensity of the sample to that of external controls. WDSV RNA/DNA ratios, based on those quantitative estimates, were computed for each organ. An RNA/DNA ratio of 3 was arbitrarily chosen as the threshold above which there was viral transcriptional activity. Viral DNA was found in all the organs examined from the three tumor-free walleyes. In those three tumor-free walleyes, low levels of WDSV RNA were detected in only one kidney and two spleen samples. In the three tumor-bearing walleyes, viral DNA was found in one brain, one kidney, two liver, and two skin samples. In contrast to the few organs from tumor-free walleyes in which WDSV RNA was detected, in tumor-bearing walleyes WDSV RNA was present in the one brain examined and in 2/3 kidney, 2/3 liver, 3/3 skin, and 3/3 spleen samples. A WDSV RNA/DNA ratio above 3 was obtained in all three tumor-bearing walleyes but in only one tumor-free fish. These data indicated that 1) both tumor-bearing and tumor-free walleyes were infected by WDSV, 2) many cell types were targeted by WDSV and supported viral transcription, and 3) tumor-bearing walleyes harbored a transcriptionally active WDSV, whereas tumor-free walleyes contained mostly silent WDSV DNA.

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