Replication of Tomato bushy stunt virus RNA in a plant in vitro system.

Abstract:

:An ideal system to investigate individual determinants of the replication process of (+)-strand RNA viruses is a cell-free extract that supports viral protein and RNA synthesis in a synchronized manner. Here, we applied a translation/replication system based on cytoplasmic extracts of Nicotiana tabacum cells to Tomato bushy stunt virus (TBSV) RNA. In vitro translated TBSV proteins p33 and p92 form viral replicase, which, in the same reaction, accomplishes the entire replication cycle on exogenous TBSV DI or full-length RNA. Tests of mutant TBSV RNAs confirmed the template specificity of the in vitro replication reaction. Complementation experiments ascertained the significance of an earlier identified TBSV host factor. Interestingly, formation of the viral replicase occurs also in the absence of concurrent protein synthesis demonstrating that translation and RNA replication are not functionally linked in this system. Our studies with cell-free extracts of a plant host thus confirmed earlier findings and enabled novel insights into the TBSV RNA replication process.

journal_name

Virology

journal_title

Virology

authors

Gursinsky T,Schulz B,Behrens SE

doi

10.1016/j.virol.2009.05.009

subject

Has Abstract

pub_date

2009-08-01 00:00:00

pages

250-60

issue

2

eissn

0042-6822

issn

1096-0341

pii

S0042-6822(09)00311-0

journal_volume

390

pub_type

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