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PMID: 6473106 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Viroid replication: equilibrium association constant and comparative activity measurements for the viroid-polymerase interaction.

Nucleic acids research ·Vol. 12 ·No. 15 ·1984-08-10 ·Pages 6231-46

Goodman TC, Nagel L, Rappold W, Klotz G, Riesner D

Abstract

The binding and replication of purified potato spindle tuber viroid (PSTV) by DNA-dependent RNA polymerase II from wheat germ was studied in analytical ultracentrifugation experiments and in vitro transcription assays. The equilibrium association constant for the viroid-polymerase interaction is 1.9 X 10(7) M-1. Both ultraviolet and fluorescent monitoring during the sedimentation experiments showed two distinguishable viroid-polymerase complexes. These are interpreted as resulting from a 1:1 and 2:1 enzyme-to-viroid binding stoichiometry. A265/A280 ratios across the sedimenting boundaries, the sedimentation velocity of the complexes, as well as electron microscopic data support this interpretation. The role of viroid secondary structure in enzyme binding and polymerization is discussed in the light of these results and compared with binding and polymerization data for virusoid RNA, single- and double-stranded RNA, and double-stranded DNA.

MeSH Terms
Microscopy, Electron Protein Binding RNA Polymerase II/genetics RNA, Viral/genetics Substrate Specificity Transcription, Genetic Viroids/genetics Virus Replication
Chemicals
RNA, Viral RNA Polymerase II
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Goodman T C
Nagel L
Rappold W
Klotz G
Riesner D
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23 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/12.15.6231
Published
1984-08-10
Pages
6231-46
Language
English
Region
England
NLM ID
0411011
PMCID
PMC320069
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/12.15.6231 PMC Full Text View journal details
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