| RRC ID |
38559
|
| Author |
Kobayashi T, Machida K, Mikami S, Masutani M, Imataka H.
|
| Title |
Cell-free RNA replication systems based on a human cell extracts-derived in vitro translation system with the encephalomyocarditisvirus RNA.
|
| Journal |
J Biochem
|
| Abstract |
To study the relationship between translation and replication of encephalomyocarditisvirus (EMCV) RNA, we established a cell-free RNA replication system by employing a human cell extracts-based in vitro translation system. In this system, a cis-EMCV RNA replicon encoding the Renilla luciferase (R-luc) or GFP and the viral regulatory proteins efficiently replicated with simultaneous translation of the encoded protein. To examine how translation of the replicon RNA, but not the translated products, affected replication, a trans-EMCV RNA replicon encoding R-luc and the RNA replication elements was next constructed. The trans-replicon RNA replicated only in the presence of the regulatory proteins pre-expressed in trans. Incubation with cycloheximide, puromycin or a dominant-negative eukaryotic translation initiation factor 4A following expression of the regulatory proteins almost completely inhibited not only translation of the trans-replicon RNA but also replication of the RNA, suggesting that EMCV RNA translation promotes replication of the RNA. In conclusion, the cell-free RNA replication systems should become useful tools for the study of the viral RNA replication.
|
| Volume |
150(4)
|
| Pages |
423-30
|
| Published |
2011-10-1
|
| DOI |
10.1093/jb/mvr072
|
| PII |
mvr072
|
| PMID |
21622667
|
| MeSH |
Animals
Cell Extracts / pharmacology*
Cell-Free System
Cells, Cultured
Cricetinae
Cycloheximide / pharmacology
Encephalomyocarditis virus / drug effects*
Encephalomyocarditis virus / genetics*
Encephalomyocarditis virus / metabolism
HeLa Cells
Humans
Puromycin / pharmacology
RNA, Viral / biosynthesis*
RNA, Viral / genetics
Virus Replication / drug effects*
Virus Replication / genetics
|
| IF |
2.476
|
| Times Cited |
6
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Human and Animal Cells |
HeLa S3 |
