High-resolution structure of an archaeal zinc ribbon defines a general architectural motif in eukaryotic RNA polymerases

AbstractBackground: Transcriptional initiation and elongation provide control points in gene expression. Eukaryotic RNA polymerase II subunit 9 (RPB9) regulates start-site selection and elongational arrest. RPB9 contains Cys4 Zn2+-binding motifs which are conserved in archaea and homologous to those of the general transcription factors TFIIB and TFIIS.Results: The structure of an RPB9 domain from the hyperthermophilic archaeon Thermococcus celer was determined at high resolution by NMR spectroscopy. The structure consists of an apical tetrahedral Zn2+-binding site, central β sheet and disordered loop. Although the structure lacks a globular hydrophobic core, the two surfaces of the β sheet each contain well ordered aromatic rings engaged in serial edge-to-face interactions. Basic sidechains are clustered near the Zn2+-binding site. The disordered loop contains sidechains conserved in TFIIS, including acidic residues essential for the stimulation of transcriptional elongation.Conclusions: The planar architecture of the RPB9 zinc ribbon – distinct from that of a conventional globular domain – can accommodate significant differences in the alignment of polar, non-polar and charged sidechains. Such divergence is associated with local and non-local changes in structure. The RPB9 structure is distinguished by a fourth β strand (extending the central β sheet) in a well ordered N-terminal segment and also differs from TFIIS (but not TFIIB) in the orientation of its apical Zn2+-binding site. Cys4 Zn2+-binding sites with distinct patterns of polar, non-polar and charged residues are conserved among unrelated RNAP subunits and predicted to form variant zinc ribbons