Simultaneous presence of terminal adenylyl, cytidylyl, guanylyl, and uridylyl transferase in healthy tomato leaf tissue: separation from RNA-dependent RNA polymerase and characterization of the terminal transferases

1982 ◽  
Vol 2 (6) ◽  
pp. 379-389 ◽  
Author(s):  
Frank Boege
Keyword(s):  
Rna Polymerase ◽  
Leaf Tissue ◽  
Tomato Leaf ◽  
Simultaneous Presence ◽  
Rna Dependent Rna Polymerase ◽  
Tissue Separation

In vitro transcription of viroid RNA into full-length copies by RNA-dependent RNA polymerase from healthy tomato leaf tissue

1982 ◽  
Vol 2 (3) ◽  
pp. 185-194 ◽  
Author(s):  
Frank Boege ◽  
Wolfgang Rohde ◽  
Heinz L. Sänger
Keyword(s):  
Rna Polymerase ◽  
Tomato Plant ◽  
Leaf Tissue ◽  
Potato Spindle Tuber Viroid ◽  
In Vitro Transcription ◽  
Tomato Leaf ◽  
Full Length ◽  
Rna Dependent Rna Polymerase ◽  
In Vitro Synthesis

RNA-dependent RNA polymerase from healthy tomato plant tissue accepts potato spindle tuber viroid (PSTV) RNA as a template for the in vitro synthesis of full-length RNA copies of the PSTV genome. Viroid transcription requires the presence of Mn2+ and/or Mg2+ ions and is not inhibited by concentrations of 10−5 M α-amanitin. This is the first report of a well-defined product synthesized in vitro by an RNA-dependent RNA polymerase from healthy plants.

scholarly journals RNA-dependent RNA polymerase from healthy tomato leaf tissue

FEBS Letters ◽  
1980 ◽  
Vol 121 (1) ◽  
pp. 91-96 ◽  
Author(s):  
Frank Boege ◽  
L.Sänger Heinz
Keyword(s):  
Rna Polymerase ◽  
Leaf Tissue ◽  
Tomato Leaf ◽  
Rna Dependent Rna Polymerase

scholarly journals Analysis of Ribonucleotide 5′-Triphosphate Analogs as Potential Inhibitors of Zika Virus RNA-Dependent RNA Polymerase by Using Nonradioactive Polymerase Assays

2016 ◽  
Vol 61 (3) ◽  
Author(s):  
Gaofei Lu ◽  
Gregory R. Bluemling ◽  
Paul Collop ◽  
Michael Hager ◽  
Damien Kuiper ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Zika Virus ◽  
Nonstructural Protein ◽  
Rna Dependent Rna Polymerase ◽  
Antiviral Compounds ◽  
Double Stranded Dna ◽  
Virus Rna ◽  
Potential Inhibitors

ABSTRACT Zika virus (ZIKV) is an emerging human pathogen that is spreading rapidly through the Americas and has been linked to the development of microcephaly and to a dramatically increased number of Guillain-Barré syndrome cases. Currently, no vaccine or therapeutic options for the prevention or treatment of ZIKV infections exist. In the study described in this report, we expressed, purified, and characterized full-length nonstructural protein 5 (NS5) and the NS5 polymerase domain (NS5pol) of ZIKV RNA-dependent RNA polymerase. Using purified NS5, we developed an in vitro nonradioactive primer extension assay employing a fluorescently labeled primer-template pair. Both purified NS5 and NS5pol can carry out in vitro RNA-dependent RNA synthesis in this assay. Our results show that Mn2+ is required for enzymatic activity, while Mg2+ is not. We found that ZIKV NS5 can utilize single-stranded DNA but not double-stranded DNA as a template or a primer to synthesize RNA. The assay was used to compare the efficiency of incorporation of analog 5′-triphosphates by the ZIKV polymerase and to calculate their discrimination versus that of natural ribonucleotide triphosphates (rNTPs). The 50% inhibitory concentrations for analog rNTPs were determined in an alternative nonradioactive coupled-enzyme assay. We determined that, in general, 2′-C-methyl- and 2′-C-ethynyl-substituted analog 5′-triphosphates were efficiently incorporated by the ZIKV polymerase and were also efficient chain terminators. Derivatives of these molecules may serve as potential antiviral compounds to be developed to combat ZIKV infection. This report provides the first characterization of ZIKV polymerase and demonstrates the utility of in vitro polymerase assays in the identification of potential ZIKV inhibitors.

Biochemical Characterization of Respiratory Syncytial Virus RNA-Dependent RNA Polymerase Complex

2020 ◽  
Vol 6 (10) ◽  
pp. 2800-2811
Author(s):  
Anand Balakrishnan ◽  
Edmund Price ◽  
Catherine Luu ◽  
Jacob Shaul ◽  
Charles Wartchow ◽  
...  
Keyword(s):  
Respiratory Syncytial Virus ◽  
Rna Polymerase ◽  
Biochemical Characterization ◽  
Rna Dependent Rna Polymerase ◽  
Polymerase Complex ◽  
Virus Rna ◽  
Syncytial Virus

The Use of Synthetic Oligo-RNA and -DNA as Defined Templates for the Determination Of Catalytic Properties of RNA-Directed RNA Polymerase from Tomato Leaf Tissue

1988 ◽  
Vol 7 (5-6) ◽  
pp. 713-716
Author(s):  
Bernd Haas ◽  
Winfried Schiebel ◽  
Snezan Marinkovic ◽  
Andrea Klanner ◽  
Heinz L. Sanger
Keyword(s):  
Rna Polymerase ◽  
Catalytic Properties ◽  
Leaf Tissue ◽  
Tomato Leaf ◽  
Rna And Dna

Production and characterization of active hepatitis C virus RNA-dependent RNA polymerase

2010 ◽  
Vol 71 (2) ◽  
pp. 147-152 ◽  
Author(s):  
Kisun Ryu ◽  
Kyun-Hwan Kim ◽  
Seong-Yeon Yoo ◽  
Eun-Young Lee ◽  
Keo-Heun Lim ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Rna Polymerase ◽  
Hepatitis C Virus Rna ◽  
Rna Dependent Rna Polymerase ◽  
Virus Rna
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