Geminiviruses employ host DNA glycosylases to subvert DNA methylation-mediated defense.

DNA methylation is an epigenetic mechanism that plays important roles in gene regulation and transposon silencing. Active DNA demethylation has evolved to counterbalance DNA methylation at many endogenous loci. Here, we report that active DNA demethylation also targets viral DNAs, tomato yellow leaf curl China virus (TYLCCNV) and its satellite tomato yellow leaf curl China betasatellite (TYLCCNB), to promote their virulence. We demonstrate that the ßC1 protein, encoded by TYLCCNB, interacts with a ROS1-like DNA glycosylase in Nicotiana benthamiana and with the DEMETER (DME) DNA glycosylase in Arabidopsis thaliana. The interaction between ßC1 and DME facilitates the DNA glycosylase activity to decrease viral DNA methylation and promote viral virulence. These findings reveal that active DNA demethylation can be regulated by a viral protein to subvert DNA methylation-mediated defense.

ßC1 protein encoded in geminivirus satellite concertedly targets MKK2 and MPK4 to counter host defense.

Plant viruses have evolved multiple strategies to overcome host defense to establish an infection. Here, we identified two components of a host mitogen-activated protein kinase (MAPK) cascade, MKK2 and MPK4, as bona fide targets of the ßC1 protein encoded by the betasatellite of tomato yellow leaf curl China virus (TYLCCNV). ßC1 interacts with the kinase domain of MKK2 and inhibits its activity. In vivo, ßC1 suppresses flagellin-induced MAPK activation and downstream responses by targeting MKK2. Furthermore, ßC1 also interacts with MPK4 and inhibits its kinase activity. TYLCCNV infection induces the activation of the MAPK cascade, mutation in MKK2 or MPK4 renders the plant more susceptible to TYLCCNV, and can complement the lack of ßC1. This work shows for the first time that a plant virus both activates and suppresses a MAPK cascade, and the discovery of the ability of ßC1 to selectively interfere with the host MAPK activation illustrates a novel virulence function and counter-host defense mechanism of geminiviruses.