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| Protein-RNA interactions play a key role in many fundamental processes and can be found in large complexes in the cell. Visualization by transmission electron microscopy (TEM) provides structural information at a resolution that is difficult to obtain in any other way. Yet, localization and tracing by electron microscopy of RNA within such large biological assemblies, are not yet a straightforward undertaking. Visualization of nucleic acid molecules by TEM cannot be directly achieved because of the low-density weakly scattering atoms they contain. Tagging such macromolecules with clusters of heavy atoms should facilitate their visualization by conventional TEM. The present invention teaches a general systematic strategy for incorporating gold clusters into nucleotides and nucleic acid molecules, thereby enabling their direct visualization by microscopical methods. This method may help in elucidating the mode of action of RNA-protein complexes.Protein-RNA interactions play key role fundamental life pathway gene expression transcribing complexes polymerase DNA template precursor messenger (pre-mRNA) associated multitude proteins small molecules large ribonucleoprotein (RNP) spliceosome mature synthesis cytoplasm ribosomes inherently polymorphic assemblies visualization transmission electron microscopy (TEM) structural information resolution difficult Yet localization tracing ribonucleotides biological yet straightforward undertaking Even crystals amenable X-ray crystallography analysis case demand heavy atom derivatives phase diffraction data Covalent derivatization atoms ensure introduction electron-dense centers distinct locations crystallized |
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