|Professor in the Department of Biosciences||CG246 (Chemistry)|
|Visiting Associate Professor in the Department of Chemistry||CG236|
|Member of the Biophysical Sciences Institute|
Toxin-antitoxin systems and bacteriophage-resistance
Whilst bacteria are often thought of as selfish cells working for their own benefit, we can observe that they exist as diverse interacting communities. This is reflected in the ubiquitous presence and implementation of "toxin-antitoxin" systems throughout known Bacterial and Archaeal species. Toxin-antitoxin systems are characterised as small genetic loci generally encoding two parts. The toxin, when free to act, will target the host cell and stall growth. In the presence of the antitoxin, this effect is negated and cells grow freely.
It might appear peculiar that bacterial cells carry toxin-antitoxin systems, until you consider the potential advantages. For instance, if there aren't enough nutrients to go around, one cell activates its internal toxins, allowing it to grow slower or die, so that the population of clonal bacteria around it can survive. Another example would be when a bacterial cell becomes infected by a bacteria-specific virus, called a bacteriophage (phage). Unchecked, the phage would replicate, burst out, and infect neighbour cells. If the infected cell shuts down quickly, however, it can stop viral spread and protect the bacterial population.
Toxins from toxin-antitoxin systems often target the same biomolecules as antibiotics. Studying how these toxins kill bacteria will allow us to develop new ideas and methods for stopping bacterial infections.
Figure 1. The BrxU DNA-modification dependent restriction enzyme targets modifed phage DNA for cleavage. This crystal structure shows an intertwined BrxU dimer, with each monomer coloured in cyans or pinks. See Picton et al. 2021, Nucleic Acids Research 49(19): 11257-11273.
Harnessing molecular tools from phage-host interactions
The interactions between phages and their bacterial hosts have generated a wealth of tools used in biotechnology, including CRISPR-Cas systems and restriction enzymes (Figure. 1).
As the natural predators of bacteria, it is also essential to investigate phage-host interactions in order to develop phages as a viable therapeutic alternative to antibiotics.
We investigate toxin-antitoxin systems and phage-host interactions using a range of molecular biology, microbiology, genetic and biochemical techniques. These include protein biochemistry, genomics and structural analysis through X-ray crystallography.
- Molecular Microbiology
- Antimicrobial Resistance
- Bacteriophage biology
- Structural Biology
- Toxin-antitoxin systems
- Phage defence
- Bandak, A. F., Blower, T. R., Nitiss, K. C., Gupta, R., Lau, A. Y., Guha, R., Nitiss, J. L. & Berger, J. M. (2023). Naturally mutagenic sequence diversity in a human type II topoisomerase. Proceedings of the National Academy of Sciences 120(28): e2302064120.
- Macdonald, E., Wright, R., Connolly, J. P. R., Strahl, H., Brockhurst, M., van Houte, S., Blower, T. R., Palmer, T. & Mariano, G. (2023). The novel anti-phage system Shield co-opts an RmuC domain to mediate phage defense across Pseudomonas species. PLOS Genetics 19(6): e1010784.
- Kelly, A., Arrowsmith, T.J., Went, S.C. & Blower, T.R. (2023). Toxin–antitoxin systems as mediators of phage defence and the implications for abortive infection. Current Opinion in Microbiology 73: 102293.
- Mariano, G. & Blower, T.R. (2023). Conserved domains can be found across distinct phage defence systems. Molecular Microbiology
- Guillen-Garcia, A., Gibson, S., Jordan, C., Ramaswamy, V., Linthwaite, V., Bromley, E.H.C., Brown, A.P. Hodgson, D., Blower, T., Verlet, J., Degiacomi, M., Palsson, L-O. & Cann, M.J. (2022). Allophycocyanin A is a carbon dioxide receptor in the cyanobacterial phycobilisome. Nature Communications 13: 5289.
- Beck, I. N., Picton, D. M. & Blower, T. R. (2022). Crystal structure of the BREX phage defence protein BrxA. Current Research in Structural Biology
- Ling, E. M., Baslé, A., Cowell, I. G., Van Den Berg, B., Blower, T. R. & Austin, C. A. (2022). A comprehensive structural analysis of the ATPase domain of Human DNA topoisomerase II Beta bound to AMPPNP, ADP and the bisdioxopiperazine, ICRF193. Structure 30(8): P1129-1145.e3.
- Picton, D. M., Harling-Lee, J. D., Duffner, S. J., Went, S. C., Morgan, R. D., Hinton, J. C. D. & Blower, T. R. (2022). A widespread family of WYL-domain transcriptional regulators co-localises with diverse phage defence systems and islands. Nucleic Acids Research 50(9): 5191-5207.
- Picton, D.M., Luyten, Y.A., Morgan, R.D., Nelson, A., Smith, D.L., Dryden, D.T.F., Hinton, J.C.D. & Blower, T.R. (2021). The phage defence island of a multidrug resistant plasmid uses both BREX and type IV restriction for complementary protection from viruses. Nucleic Acids Research 49(19): 11257-11273.
- Usher, B., Birkholz, N., Beck, I.N., Fagerlund, R.D., Jackson, S.A., Fineran, P.C. & Blower, T.R. (2021). Crystal structure of the anti-CRISPR repressor Aca2. Journal of Structural Biology 213(3): 107752.
- Rodwell, E.V., Wenner, N., Pulford, C.V., Cai, Y., Bowers-Barnard, A., Beckett, A., Rigby, J., Picton, D.M., Blower, T.R., Feasey, N.A., Hinton, J.C.D. & Perez-Sepulveda, B.M. (2021). Isolation and characterisation of bacteriophages with activity against invasive non-typhoidal Salmonella causing blood-stream infection in Malawi. Viruses 13(3): 478.
- Cai, Y., Usher, B., Gutierrez, C., Tolcan, A., Mansour, M., Fineran, P.C., Condon, C., Neyrolles, O., Genevaux, P. & Blower, T.R. (2020). A nucleotidyltransferase toxin inhibits growth of Mycobacterium tuberculosis through inactivation of tRNA acceptor stems. Science Advances 6(31): eabb6651.
- Beck, I.N., Usher, B., Hampton, H.G., Fineran, P.C. & Blower, T.R. (2020). Antitoxin autoregulation of M. tuberculosis toxin-antitoxin expression through negative cooperativity arising from multiple inverted repeat sequences. Biochemical Journal 477(12): 2401-2419.
- Cross, J.M., Blower, T.R., Kingdon, A.D.H., Pal, R., Picton, D.M. & Walton, J.W. (2020). Anticancer Ruthenium Complexes with HDAC Isoform Selectivity. Molecules 25(10): 2383.
- Blower, T.R., Bandak, A., Lee, A.S.Y., Austin, C.A., Nitiss, J.L. & Berger, J.M. (2019). A complex suite of loci and elements in eukaryotic type II topoisomerases determine selective sensitivity to distinct poisoning agents. Nucleic Acids Research 47(15): 8163-8179.
- Gibson, E.G., Blower, T.R., Cacho, M., Bax, B., Berger, J.M. & Osheroff, N. (2018). Mechanism of Action of Mycobacterium tuberculosis Gyrase Inhibitors: A Novel Class of Gyrase Poisons. ACS Infectious Diseases 4(8): 1211-1222.
- Hampton, H.G., Jackson, S.A., Fagerlund, R.D., Vogel, A.I.M., Dy, R.L., Blower, T.R. & Fineran, P.C. (2018). AbiEi binds cooperatively to the Type IV abiE toxin-antitoxin operator via a positively-charged surface and causes DNA bending and negative autoregulation. Journal of Molecular Biology 430(8): 1141-1156.
- Ashley, R.E., Blower, T.R., Berger, J.M. & Osheroff, N. (2017). Recognition of DNA Supercoil Geometry by Mycobacterium tuberculosis Gyrase. Biochemistry 56(40): 5440-5448.
- Blower, T.R., Chai, R., Przybilski, R., Chindhy, S., Fang, X., Kidman, S.E., Tan, H., Luisi, B.F., Fineran, P.C. & Salmond, G.P.C. (2017). Evolution of Pectobacterium bacteriophage ΦM1 to escape two bifunctional Type III toxin-antitoxin and abortive infection systems through mutations in a single viral gene. Applied and Environmental Microbiology 83(8): e03229-16.
- Cross, J.M., Blower, T.R., Gallagher, N., Gill, J.H., Rockley, K.L. & Walton, J.W. (2016). Anticancer RuII and RhIII Piano-Stool Complexes that are Histone Deacetylase Inhibitors. ChemPlusChem 81(12): 1276-1280.
- Blower, T.R., Williamson, B.H., Kerns, R.J. & Berger, J.M. (2016). Crystal structure and stability of gyrase–fluoroquinolone cleaved complexes from Mycobacterium tuberculosis. Proceedings of the National Academy of Sciences 113(7): 1706-1713.
- Aldred, K.J., Blower, T.R., Kerns, R.J., Berger, J.M. & Osheroff, N. (2016). Fluoroquinolone interactions with Mycobacterium tuberculosis gyrase: Enhancing drug activity against wild-type and resistant gyrase. Proceedings of the National Academy of Sciences 113(7): E839-E846.
- Rao, F., Short, F.L., Voss, J.E., Blower, T.R., Orme, A.L., Whittaker, T.E., Luisi, B.F. & Salmond, G.P.C. (2015). Co-evolution of quaternary organization and novel RNA tertiary interactions revealed in the crystal structure of a bacterial protein–RNA toxin–antitoxin system. Nucleic Acids Research 43(19): 9529-9540.
- Unwin, R.R., Cabanas, R.A., Yanagishima, T., Blower, T.R., Takahashi, H., Salmond, G.P.C., Edwardson, J.M., Fraden, S. & Eiser, E. (2015). DNA driven self-assembly of micron-sized rods using DNA-grafted bacteriophage fd virions. Physical chemistry chemical physics 17(12): 8194-202.
- Short, F.L., Pei, X.Y., Blower, T.R., Ong, S.L., Fineran, P.C., Luisi, B.F. & Salmond, G.P. (2013). Selectivity and self-assembly in the control of a bacterial toxin by an antitoxic noncoding RNA pseudoknot. Proceedings of the National Academy of Sciences of the United States of America 110(3): E241-9.
- Blower, T.R., Short, F.L., Fineran, P.C. & Salmond, G.P. (2012). Viral molecular mimicry circumvents abortive infection and suppresses bacterial suicide to make hosts permissive for replication. Bacteriophage 2(4): 234-238.
- Blower, T.R., Evans, T.J., Przybilski, R., Fineran, P.C. & Salmond, G.P. (2012). Viral Evasion of a Bacterial Suicide System by RNA-Based Molecular Mimicry Enables Infectious Altruism. PLoS Genetics 8(10): e1003023.
- Short, F.L., Blower, T.R. & Salmond, G.P. (2012). A promiscuous antitoxin of bacteriophage T4 ensures successful viral replication. Molecular Microbiology 83(4): 665-668.
- Blower, T.R., Short, F.L., Rao, F., Mizuguchi, K., Pei, X.Y., Fineran, P.C., Luisi, B.F. & Salmond, G.P. (2012). Identification and classification of bacterial Type III toxin-antitoxin systems encoded in chromosomal and plasmid genomes. Nucleic Acids Research 40(13): 6158-6173.
- Blower, T.R., Salmond, G.P.C. & Luisi, B.F. (2011). Balancing at survival's edge: the structure and adaptive benefits of prokaryotic toxin-antitoxin partners. Current Opinion in Structural Biology 21(1): 109-18.
- Blower, T.R., Pei, X.Y., Short, F.L., Fineran, P.C., Humphreys, D.P., Luisi, B.F. & Salmond, G.P.C. (2011). A processed non-coding RNA regulates an altruistic bacterial antiviral system. Nature Structural and Molecular Biology 18(2): 185-190.
- Blower, T.R., Fineran, P.C., Johnson, M.J., Toth, I.K., Humphreys, D.P. & Salmond, G.P. (2009). Mutagenesis and functional characterisation of the RNA and protein components of the toxIN abortive infection and toxin-antitoxin locus of Erwinia. Journal of Bacteriology 191(19): 6029-6039.
- Fineran, P.C., Blower, T.R., Foulds, I.J., Humphreys, D.P., Lilley, K.S. & Salmond, G.P. (2009). The phage abortive infection system, ToxIN, functions as a protein-RNA toxin-antitoxin pair. Proceedings of the National Academy of Sciences of the United States of America 106(3): 894-899.