biostudies-literatureChen YNIAID NIH HHSNIGMS NIH HHS12544-55https://www.ebi.ac.uk/biostudies/studies/S-EPMC3154994biostudies-literatureUnknown133(32)The nucleotidyl transfer reaction leading to formation of the first phosphodiester bond has been followed in real time by Raman microscopy, as it proceeds in single crystals of the N4 phage virion RNA polymerase (RNAP). The reaction is initiated by soaking nucleoside triphosphate (NTP) substrates and divalent cations into the RNAP and promoter DNA complex crystal, where the phosphodiester bond formation is completed in about 40 min. This slow reaction allowed us to monitor the changes of the RNAP and DNA conformations as well as bindings of substrate and metal through Raman spectra taken every 5 min. Recently published snapshot X-ray crystal structures along the same reaction pathway assisted the spectroscopic assignments of changes in the enzyme and DNA, while isotopically labeled NTP subJournal of the American Chemical SocietyTime-resolved events on the reaction pathway of transcript initiation by a single-subunit RNA polymerase: Raman crystallographic evidence.PMC3154994R01 GM071897R01 GM081420R01 GM081420-04GM54072R56 GM054072GM81420GM071897AI12575R01 GM054072R01 GM071897-04R01 AI012575R56 GM054072-13Carey PRGleghorn MLMurakami KSChen YBasu RfalseTime-resolved events on the reaction pathway of transcript initiation by a single-subunit RNA polymerase: Raman crystallographic evidence.The nucleotidyl transfer reaction leading to formation of the first phosphodiester bond has been followed in real time by Raman microscopy, as it proceeds in single crystals of the N4 phage virion RNA polymerase (RNAP). The reaction is initiated by soaking nucleoside triphosphate (NTP) substrates and divalent cations into the RNAP and promoter DNA complex crystal, where the phosphodiester bond formation is completed in about 40 min. This slow reaction allowed us to monitor the changes of the RNAP and DNA conformations as well as bindings of substrate and metal through Raman spectra taken every 5 min. Recently published snapshot X-ray crystal structures along the same reaction pathway assisted the spectroscopic assignments of changes in the enzyme and DNA, while isotopically labeled NTP sub2011-01-01T00:00:00Z2011 Aug2025-05-29T21:53:01.851Z2025-05-29T21:53:01.851ZS-EPMC31549942174480610.1021/ja201557w