Abstract
Riboswitches constitute an important class of non coding RNAs, present in the UTR of mRNAs. The ligand discriminating aptamer domain of a riboswitch can undergo conformational changes in absence or presence of its binding cognate metabolite affecting the downstream expression platform, through the activation or attenuation of either transcription or translation. The preQ1 responsive family of riboswitches, which show high affinity towards the guanine derived metabolites such as 7-cyano-7-deazaguanine (PreQ0) and 7-aminomethyl-7-deazaguanine (PreQ1), consist of the smallest riboswitches known. There are two subclasses of preQ1 riboswitches categorised based on sequence and structural differences: Class I and Class II. Class I is further subdivided into Type I and Type II, based on the mechanism of gene regulation. Class I Type I riboswitches regulate gene expression at translational level, and Class I Type II riboswitches regulate gene expression at transcriptional level. X-ray crystal structures of aptamer domains of Class I Types I and II riboswitches bound with cognate ligands preQ0 and preQ1 have been reported. However, the ligand free structure of only Class I Type I is available. We have modelled both ligand bound and ligand free structures of Class I Type I riboswitches belonging to Thermoanaerobacter tengcongensis. Crystal structures show that the presence or absence of ligand results in changes in molecular interactions within the binding pocket, eventually resulting in either sequestration or opening of ribosomal binding site leading to translational regulation. In order to study the stability of base pairs in the aptamer region, we have modelled the interactions observed in binding pocket, both pairing and stacking, and carried out gas phase quantum chemical studies. The ideal geometries and interaction energies of the complexes, evaluated at B3LYP/6-31G(d,p)//RIMP2/aug-cc-pVDZ respectively. The effect of stacking interactions in the complex is evaluated with DFT-D calculations at PBE/6-31G(d,p) level of theory. Morokuma energy decomposition is performed to investigate the contributions of various non covalent interactions to the total energy of the complex. We have identified the contributions of different interactions towards the stability of binding pocket and recognition of the ligands. Our studies also provide insights into the differential ligand affinities of the aptamer domain towards two similar ligands: preQ0 and preQ1.