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Title: Transcription of potato spindle tuber viroid by RNA polymerase II starts in the left terminal loop

Abstract

Viroids are single-stranded, circular RNAs of 250 to 400 bases, that replicate autonomously in their host plants but do not code for a protein. Viroids of the family Pospiviroidae, of which potato spindle tuber viroid (PSTVd) is the type strain, are replicated by the host's DNA-dependent RNA polymerase II in the nucleus. To analyze the initiation site of transcription from the (+)-stranded circles into (-)-stranded replication intermediates, we used a nuclear extract from a non-infected cell culture of the host plant S. tuberosum. The (-)-strands, which were de novo-synthesized in the extract upon addition of circular (+)-PSTVd, were purified by affinity chromatography. This purification avoided contamination by host nucleic acids that had resulted in a misassignment of the start site in an earlier study. Primer-extension analysis of the de novo-synthesized (-)-strands revealed a single start site located in the hairpin loop of the left terminal region in circular PSTVd's secondary structure. This start site is supported further by analysis of the infectivity and replication behavior of site-directed mutants in planta.

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2]
  1. Institut fuer Physikalische Biologie, Heinrich-Heine-Universitaet Duesseldorf, Geb. 26.12.U1, Universitaetsstr.1, D-40225 Duesseldorf (Germany)
  2. Institut fuer Physikalische Biologie, Heinrich-Heine-Universitaet Duesseldorf, Geb. 26.12.U1, Universitaetsstr.1, D-40225 Duesseldorf (Germany). E-mail: riesner@biophys.uni-duesseldorf.de
Publication Date:
OSTI Identifier:
20779481
Resource Type:
Journal Article
Resource Relation:
Journal Name: Virology; Journal Volume: 347; Journal Issue: 2; Other Information: DOI: 10.1016/j.virol.2005.11.039; PII: S0042-6822(05)00789-0; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CELL CULTURES; CHROMATOGRAPHY; DNA; HOST; INFECTIVITY; MUTANTS; POTATOES; PROTEINS; RNA; TRANSCRIPTION

Citation Formats

Kolonko, Nadine, Bannach, Oliver, Aschermann, Katja, Hu, Kang-Hong, Moors, Michaela, Schmitz, Michael, Steger, Gerhard, and Riesner, Detlev. Transcription of potato spindle tuber viroid by RNA polymerase II starts in the left terminal loop. United States: N. p., 2006. Web. doi:10.1016/J.VIROL.2005.1.
Kolonko, Nadine, Bannach, Oliver, Aschermann, Katja, Hu, Kang-Hong, Moors, Michaela, Schmitz, Michael, Steger, Gerhard, & Riesner, Detlev. Transcription of potato spindle tuber viroid by RNA polymerase II starts in the left terminal loop. United States. doi:10.1016/J.VIROL.2005.1.
Kolonko, Nadine, Bannach, Oliver, Aschermann, Katja, Hu, Kang-Hong, Moors, Michaela, Schmitz, Michael, Steger, Gerhard, and Riesner, Detlev. Mon . "Transcription of potato spindle tuber viroid by RNA polymerase II starts in the left terminal loop". United States. doi:10.1016/J.VIROL.2005.1.
@article{osti_20779481,
title = {Transcription of potato spindle tuber viroid by RNA polymerase II starts in the left terminal loop},
author = {Kolonko, Nadine and Bannach, Oliver and Aschermann, Katja and Hu, Kang-Hong and Moors, Michaela and Schmitz, Michael and Steger, Gerhard and Riesner, Detlev},
abstractNote = {Viroids are single-stranded, circular RNAs of 250 to 400 bases, that replicate autonomously in their host plants but do not code for a protein. Viroids of the family Pospiviroidae, of which potato spindle tuber viroid (PSTVd) is the type strain, are replicated by the host's DNA-dependent RNA polymerase II in the nucleus. To analyze the initiation site of transcription from the (+)-stranded circles into (-)-stranded replication intermediates, we used a nuclear extract from a non-infected cell culture of the host plant S. tuberosum. The (-)-strands, which were de novo-synthesized in the extract upon addition of circular (+)-PSTVd, were purified by affinity chromatography. This purification avoided contamination by host nucleic acids that had resulted in a misassignment of the start site in an earlier study. Primer-extension analysis of the de novo-synthesized (-)-strands revealed a single start site located in the hairpin loop of the left terminal region in circular PSTVd's secondary structure. This start site is supported further by analysis of the infectivity and replication behavior of site-directed mutants in planta.},
doi = {10.1016/J.VIROL.2005.1},
journal = {Virology},
number = 2,
volume = 347,
place = {United States},
year = {Mon Apr 10 00:00:00 EDT 2006},
month = {Mon Apr 10 00:00:00 EDT 2006}
}