Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization

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  • Julie A.Z. Zedler
  • Alexandra M. Schirmacher
  • David A. Russo
  • Lorna Hodgson
  • Emil Gundersen
  • Annemarie Matthes
  • Stefanie Frank
  • Paul Verkade
  • Jensen, Poul Erik

Cyanobacteria offer great potential as alternative biotechnological hosts due to their photoautotrophic capacities. However, in comparison to established heterotrophic hosts, several key aspects, such as product titers, are still lagging behind. Nanobiotechnology is an emerging field with great potential to improve existing hosts, but so far, it has barely been explored in microbial photosynthetic systems. Here, we report the establishment of large proteinaceous nanofilaments in the unicellular model cyanobacterium Synechocystis sp. PCC 6803 and the fast-growing cyanobacterial strain Synechococcus elongatus UTEX 2973. Transmission electron microscopy and electron tomography demonstrated that expression of pduA*, encoding a modified bacterial microcompartment shell protein, led to the generation of bundles of longitudinally aligned nanofilaments in S. elongatus UTEX 2973 and shorter filamentous structures in Synechocystis sp. PCC 6803. Comparative proteomics showed that PduA* was at least 50 times more abundant than the second most abundant protein in the cell and that nanofilament assembly had only a minor impact on cellular metabolism. Finally, as a proof-of-concept for co-localization with the filaments, we targeted a fluorescent reporter protein, mCitrine, to PduA* by fusion with an encapsulation peptide that natively interacts with PduA. The establishment of nanofilaments in cyanobacterial cells is an important step toward cellular organization of heterologous pathways and the establishment of cyanobacteria as next-generation hosts.

TidsskriftACS Nano
Udgave nummer24
Sider (fra-til)25279-25290
Antal sider12
StatusUdgivet - 2023

Bibliografisk note

Funding Information:
The authors thank C. Neal and J. Mantell of the Wolfson Bioimaging Facility of the University of Bristol for their expert assistance with transmission electron microscopy, M. Jones and P. Curnow for providing JAZZ access to their facilities at the University of Bristol, M. Warren and M. Lee for providing the PduA* DNA sequence, and C. Crocoll for help with data management. The authors acknowledge financial support from the “Cynthetica” project from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement no. 745959 (JAZZ), the Humboldt Foundation (DAR), BBSRC sLoLa Research grant (BB/M002969/1) (PV), the Novo Nordisk Foundation (NNF19OC0057634) (PEJ), and the Carlsberg Foundation (CF17-0657) (PEJ).

Publisher Copyright:
© 2023 The Authors. Published by American Chemical Society.

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