Detection of an en masse and reversible B- to A-DNA conformational transition in prokaryotes in response to desiccation

Abstract

The role that DNA conformation plays in the biochemistry of cells has been the subject of intensive research since DNA polymorphism was discovered. B-DNA has long been considered the native form of DNA in cells although alternative conformations of DNA are thought to occur transiently and along short tracts. Here, we report the first direct observation of a fully reversible en masse conformational transition between B- and A-DNA within live bacterial cells using Fourier transform infrared (FTIR) spectroscopy. This biospectroscopic technique allows for non-invasive and reagent-free examination of the holistic biochemistry of samples. For this reason, we have been able to observe the previously unknown conformational transition in all four species of bacteria investigated. Detection of this transition is evidence of a previously unexplored biological significance for A-DNA and highlights the need for new research into the role that A-DNA plays as a cellular defence mechanism and in stabilizing the DNA conformation. Such studies are pivotal in understanding the role of A-DNA in the evolutionary pathway of nucleic acids. Furthermore, this discovery demonstrates the exquisite capabilities of FTIR spectroscopy and opens the door for further investigations of cell biochemistry with this under-used technique.

Keywords: A-DNA; DNA conformation; attenuated total reflection–Fourier transform infrared spectroscopy; bacteria; infrared.

Figure 1.

FTIR spectra (a) and calculated second derivative spectra (b) of double-stranded DNA in the dehydrated A-DNA conformation (dashed red) and after rehydration in the B-DNA conformation (solid blue). Shading indicates, from left to right, the anti-symmetric phosphate stretch, the symmetric phosphate stretch and the backbone carbon–carbon vibration. (Online version in colour.)

Figure 2.

FTIR spectra (a) and calculated second derivatives (b) of P. vulgaris in the dormant dehydrated state (dashed red) and after rehydration (solid blue). From left to right, shading highlights the characteristic anti-symmetric phosphate stretch, the symmetric phosphate stretch and the backbone carbon–carbon vibration that are diagnostic of the A- to B-DNA transition. (Online version in colour.)

Figure 3.

FTIR spectra (a) and calculated second derivatives (b) of E. coli in the dormant dehydrated state (dashed red) and after rehydration (solid blue). From left to right, shading highlights the characteristic anti-symmetric phosphate stretch, the symmetric phosphate stretch and the backbone carbon–carbon vibration that are diagnostic of the A- to B-DNA transition. (Online version in colour.)

Figure 4.

PCA scores plot of 12 FTIR spectra of hydrated (light blue) and dehydrated (dark red) bacteria in principal component 1 and 2 space. (Online version in colour.)