Discovery of a small molecule that inhibits bacterial ribosome biogenesis

Abstract

While small molecule inhibitors of the bacterial ribosome have been instrumental in understanding protein translation, no such probes exist to study ribosome biogenesis. We screened a diverse chemical collection that included previously approved drugs for compounds that induced cold sensitive growth inhibition in the model bacterium Escherichia coli. Among the most cold sensitive was lamotrigine, an anticonvulsant drug. Lamotrigine treatment resulted in the rapid accumulation of immature 30S and 50S ribosomal subunits at 15°C. Importantly, this was not the result of translation inhibition, as lamotrigine was incapable of perturbing protein synthesis in vivo or in vitro. Spontaneous suppressor mutations blocking lamotrigine activity mapped solely to the poorly characterized domain II of translation initiation factor IF2 and prevented the binding of lamotrigine to IF2 in vitro. This work establishes lamotrigine as a widely available chemical probe of bacterial ribosome biogenesis and suggests a role for E. coli IF2 in ribosome assembly.

DOI: http://dx.doi.org/10.7554/eLife.03574.001

eLife digest

Inside cells, molecular machines called ribosomes make proteins from instructions that are provided by genes. The ribosomes themselves are made up of about 50 proteins and three RNA molecules that need to be assembled like a 3-D jigsaw. In bacteria, a group of proteins called ribosome biogenesis factors help to assemble these pieces correctly.

To study how a biological process works, scientists often look at what happens when a component is missing or not working properly. However, this approach cannot be used to study how ribosomes are made because stopping protein production entirely will kill the cell. Another approach is to use chemicals to temporarily stop or slow down a biological process, but researchers are yet to find a chemical that can do this for ribosome assembly.

To address this problem, Stokes et al. ‘screened’ 30,000 chemicals in an effort to find one or more that could affect ribosome assembly in bacteria. The screen revealed that a drug called lamotrigine—which is used to treat epilepsy and other conditions in humans—could stop the assembly of ribosomes, but did not affect the production of proteins by completed ribosomes.

The experiments also suggest that initiation factor 2, a protein that is involved in the production of other proteins, may also have a role in ribosome assembly. Another recent study found that the equivalent of initiation factor 2 in yeast acts as a quality control checkpoint during ribosome assembly, so the bacterial version may also perform a similar role.

It is also be possible that lamotrigine might be used to help develop a novel mechanistic class of antibiotics.

DOI: http://dx.doi.org/10.7554/eLife.03574.002