Designing efficient genetic code expansion in Bacillus subtilis to gain biological insights

• Published in: Nature communications Vol. 12; no. 1; p. 5429
• Main Authors: Stork, Devon A., Squyres, Georgia R., Kuru, Erkin, Gromek, Katarzyna A., Rittichier, Jonathan, Jog, Aditya, Burton, Briana M., Church, George M., Garner, Ethan C., Kunjapur, Aditya M.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 14.09.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 14/35; 14/63; 631/326/41/2173; 631/326/88; 631/92/552; 82/58; 82/80; 82/83; Amino acids; Amino Acids - chemistry; Amino Acids - genetics; Amino Acids - metabolism; Amino Acyl-tRNA Synthetases - classification; Amino Acyl-tRNA Synthetases - genetics; Amino Acyl-tRNA Synthetases - metabolism; Bacillus subtilis; Bacillus subtilis - genetics; Bacillus subtilis - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biology; Cell division; Codon; Codons; Crosslinking; Cytokinesis; Cytokinesis - genetics; E coli; Escherichia coli - genetics; Escherichia coli - metabolism; Expansion; Gene Expression Regulation, Bacterial; Genetic Code; Genome, Bacterial; Gram-positive bacteria; Humanities and Social Sciences; Industrial applications; Labeling; multidisciplinary; Protein Binding; Protein Biosynthesis; Protein Interaction Mapping; Proteins; RNA, Transfer - genetics; RNA, Transfer - metabolism; Science; Science (multidisciplinary); Stop codon; Titration
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-25691-4

Bacillus subtilis is a model gram-positive bacterium, commonly used to explore questions across bacterial cell biology and for industrial uses. To enable greater understanding and control of proteins in B. subtilis , here we report broad and efficient genetic code expansion in B. subtilis by incorporating 20 distinct non-standard amino acids within proteins using 3 different families of genetic code expansion systems and two choices of codons. We use these systems to achieve click-labelling, photo-crosslinking, and translational titration. These tools allow us to demonstrate differences between E. coli and B. subtilis stop codon suppression, validate a predicted protein-protein binding interface, and begin to interrogate properties underlying bacterial cytokinesis by precisely modulating cell division dynamics in vivo. We expect that the establishment of this simple and easily accessible chemical biology system in B. subtilis will help uncover an abundance of biological insights and aid genetic code expansion in other organisms. B. subtilis is valuable both as a model for cell biology and as an industrial organism. Here the authors use genetic code expansion to enable functional tools for exploring cell division dynamics.