Selective phosphorylation of the so-called TMV movement protein (TMV-MP) also influences the spreading of tobacco mosaic virus in some tobacco species. Phosphory-lation sites were identified by replacing putative phosphorylation sites in recombinant movement protein with alanine and glycine and subsequent determination of the absence or presence of phosphorylation using radioactive ATP in vitro . Trut-nyeva and co-workers  then replaced the identified Ser and Thr phosphorylation sites in a sequential manner with negatively charged amino acids and compared virus spreading for each mutant. While the phosphorylation of TMV-MP at one specific Ser seems to stimulate cell-to-cell movement of the virus, phosphorylation of two other sites has the opposite effect. The authors conclude that this multisite phosphorylation event may proceed in a sequential manner: First, phosphorylation leads to a spreading of the virus (a benefit for the virus), but further phosphorylation events then restrict the harmful infection (a benefit for the host plant).
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