Reconstitution and expression of mcy gene cluster in the model Cyanobacterium Synechococcus 7942 reveals a roll of MC-LR in cell division.
Yanli ZhengChunling XueHui ChenAnqi JiaLiang ZhaoJunli ZhangLixin ZhangQiang WangPublished in: The New phytologist (2023)
Cyanobacterial blooms pose a serious threat to public health due to the presence of cyanotoxins. Microcystin-LR (MC-LR) produced by Microcystis aeruginosa is the most common cyanotoxins. Due to the limitation of isolation, purification, and genetic manipulation techniques, it is difficult to study and verify in situ the biosynthetic pathways and molecular mechanisms of MC-LR. We reassembled the biosynthetic gene cluster (mcy cluster) of MC-LR in vitro by synthetic biology, designed and constructed the strong bidirectional promoter biPpsbA 2 , transformed it into Synechococcus 7942, and successfully expressed MC-LR at a level of 0.006-0.018 fg cell -1 day -1 . We found the expression of MC-LR led to abnormal cell division and the cellular filamentation, further using various methods proved that by irreversibly competing its GTP binding site, MC-LR inhibits assembly of the cell division protein FtsZ. The study represents the first reconstitution and expression of the mcy cluster and the autotrophic production of MC-LR in model cyanobacterium, which lays the foundation for resolving the microcystins (MCs) biosynthesis pathway. The discovered role of MC-LR in cell division reveals a mechanism of how blooming cyanobacteria gain a competitive edge over their non-blooming counterparts.