Precisely full length, circularizable, complementary RNA: an infectious form of potato spindle tuber viroid

The replication of many viral and subviral pathogens as well as the amplification of certain cellular genes proceeds via a rolling circle mechanism. For potato spindle tuber (PSTVd) and related viroids, the possible role of a circular (-)strand RNA as a template for synthesis of (+)strand progeny is...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1998-05, Vol.95 (11), p.6560-6565
Main Authors: Feldstein, P.A. (Center for Engineering Plants for Resistance Against Pathogens, Davis, CA.), Hu, Y, Owens, R.A
Format: Article
Language:English
Subjects:
ARN
Base Sequence
Biological Sciences
Botany
Circles
CIRCULAR RNA
Electrophoresis
EXPERIMENTACION IN VIVO
EXPERIMENTAL INFECTION
EXPERIMENTATION IN VIVO
Gels
IN VIVO EXPERIMENTATION
INFECCION EXPERIMENTAL
INFECTION EXPERIMENTALE
INFECTIVITY
Inoculum
LYCOPERSICON ESCULENTUM
MOLECULAR CONFORMATION
Molecular Sequence Data
Molecules
NEPOVIRUS
NEPOVIRUSES
NICOTIANA
NICOTIANA BENTHAMIANA
Nucleic Acid Conformation
PATHOGENICITY
Pathology
PATOGENICIDAD
PLANT VIRUSES
PLANTAS TRANSGENICAS
PLANTE TRANSGENIQUE
Plasmids
Potatoes
POUVOIR PATHOGENE
REPLICACION
REPLICATION
Ribonucleic acid
RIBOZYMES
RNA
RNA probes
RNA, Viral - chemistry
RNA, Viral - genetics
Solanum tuberosum - virology
TOMATO PLANTA MACHO VIROID
TOMATO RINGSPOT NEPOVIRUS
TRANSGENIC PLANTS
VIROIDE
VIROIDES
VIROIDS
Viroids - physiology
VIRUS DE LAS PLANTAS
VIRUS DES VEGETAUX
Virus Replication
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Description
Summary:The replication of many viral and subviral pathogens as well as the amplification of certain cellular genes proceeds via a rolling circle mechanism. For potato spindle tuber (PSTVd) and related viroids, the possible role of a circular (-)strand RNA as a template for synthesis of (+)strand progeny is unclear. Infected plants appear to contain only multimeric linear (-)strand RNAs, and attempts to initiate infection with multimeric (-)PSTVd RNAs generally have failed. To examine critically the infectivity of monomeric (-)strand viroid RNAs, we have developed a ribozyme-based expression system for the production of precisely full length (-)strand RNAs whose temini are capable of undergoing facile circularization in vitro. Mechanical inoculation of tomato seedlings with electrophoretically purified (-)PSTVd RNA led to a small fraction of plants becoming infected whereas parallel assays with an analogous tomato planta macho viroid (-)RNA resulted in a much larger fraction of infected plants. Ribozyme-mediated production of (-)PSTVd RNA in transgenic plants led to the appearance of monomeric circular (-)PSTVd RNA and large amounts of (+)PSTVd progeny. No monomeric circular (-)PSTVd RNA could be detected in naturally infected plants by using either ribonuclease protection or electrophoresis under partially denaturing conditions. Although not a component of the normal replicative pathway, precisely full length (-)PSTVd RNA appears to contain all of the structural and regulatory elements necessary for initiation of viroid replication
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.95.11.6560