Cytosine methylation inhibits replication of African cassava mosaic virus by two distinct mechanisms.
Extrachromosomally replicating viral DNA is usually free of cytosine methylation and viral templates methylated in vitro are poor substrates when used in replication assays. We have investigated the mechanism of inhibition of viral replication by DNA methylation using as a model the DNA A of African cassava mosaic virus. We have constructed two component helper systems which allow for separation of the transcriptional inhibition of viral genes necessary for replication from replication inhibition due to altered interaction between the replication complex and methylated viral DNA. Our results suggest that methylation-mediated reduction of viral replication is due to both repression mechanisms and that this provides two independent selection pressures for the maintenance of methylation-free replicons in infected cells.
Ermak
G
G
Institute of Plant Sciences, ETH-Zurich, Switzerland.
Paszkowski
U
U
Wohlmuth
M
M
Mittelsten Scheid
O
O
Paszkowski
J
J
D003596:Cytosine / Q000378:metabolism*; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; D002365:Manihot; D008745:Methylation; D009029:Mosaic Viruses / Q000502:physiology*; D010957:Plasmids; D011523:Protoplasts / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection; D014779:Virus Replication
D004279:DNA, Viral; D003596:Cytosine; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific