Alexander Shekhtman

  • Alexander Shekhtman


    The goal of Dr. Shekhtman’s laboratory is to understand RNA, RNA-ligand and RNA- protein structure-function and structure-dynamics relationships at atomic resolution in vitro and in vivo by using X-ray crystallography and NMR spectroscopy. We will explore three specific areas: (i) riboswitch RNAs; (ii) RNA pseudoknots; and (iii) microRNA, RNA-ligand and RNA-protein structures in vivo. Combined usage of X- ray and NMR extends the applicability of the structural analysis allowing the study of not only static features captured in atomic resolution structures but also the dynamic nature of various RNA conformers. The cellular environment plays a significant role in determining the overall conformation, folding topology, and stability of RNAs. We developed a new technology to study RNA structure-function relationships and RNA interaction with effector proteins directly in the eukaryotic environment. Two approaches to be explored are direct injection of labeled RNAs into Xenopus laevis oocytes for eukaryotic in-cell NMR measurements and electroporation of labeled RNA into yeast or bacteria. Both methods can be conveniently manipulated to permit the direct deposition of defined quantities of exogenous nucleic acid into the cellular environment. The major questions to be addressed by using in-cell NMR are the stability of RNA structures and structural changes upon ligand binding under physiological conditions. We are using a well-studied Pre-Q1 RNA pseudoknot to investigate the changes in secondary and tertiary structure due to intracellular interactions.