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Arabidopsis CPK3 function in anti-viral response: importance of plasma membrane nanoscale dynamics
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International audience. Viruses propagate in plant cells through the plasma membrane channels plasmodesmata. Using transient expression in tobacco leaves, we previously showed that the calcium-dependent protein kinase 3 (AtCPK3) from Arabidopsis was able to restrict viral cell-to-cell movement of the potato virus X. Importantly, AtCPK3 phosphorylated the potato Remorin StREM1.3, which was required for its anti-viral function. Using the plantago asiatica mosaic virus able to infect Arabidopsis, we now showed that this mechanism was conserved in Arabidopsis. We tested several higher order cpk knockout mutants and discovered that AtCPK3 displayed a specific ability to hamper viral propagation over 5 other CPK isoforms (AtCPK1/2/5/6/11) involved in immune response to bacteria and fungi. We confirmed that Arabidopsis AtREM1.2 was also an in vitro substrate of AtCPK3. Both AtREM1.2 and AtCPK3 are membrane-associated, which is required for their ability to restrict viral propagation. Using single particle tracking photo-activated localization microscopy, we showed that CPK3 diffusion in the plasma membrane was reduced upon activation as well as upon viral infection and that such immobilization depended on AtREM1.2. By contrast, AtREM1.2 diffusion was increased by viral infection in an AtCPK3-dependent manner. This study unveils a complex membrane protein nanoscale dynamics as part of plant defence against viruses.
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