ABSTRACT
Small acidic peptides involved in gene expression have been isolated from prokaryotic and eukaryotic cells. Synthetic peptides, designed on the basis of native peptides characteristics, show a biological activity similar to that of native peptides in in vitro reconstituted systems. These synthetic peptides are able to bind to DNA in presence of divalent cations (Cu2+, Fe2+, Mg2+) and salt/ethanol.
Subject(s)
Cations, Divalent/metabolism , DNA/metabolism , Oligopeptides/chemical synthesis , Oligopeptides/metabolism , Phosphopeptides/metabolism , Animals , Casein Kinase II , Cattle , Ethanol , Phosphorylation , Protein Serine-Threonine Kinases , RNA Polymerase II/chemical synthesis , RNA Polymerase II/metabolismABSTRACT
Peptides representing single repeat units of the carboxy-terminal domain (CTD) of RNA polymerase II (Tyr-Ser-Pro-Thr-Ser-Pro-Ser-Tyr-NH2, 1) contain overlapping Ser-Pro-Xaa-Xaa beta-turn forming sites which permit their overall structure to closely resemble members of the quinoxaline class of antitumor DNA bisintercalators. We have modified this native sequence at the i+2 positions of each beta-turn unit by substituting Gly or D-Ala in an attempt to preorganize this structure in aqueous solution. CD and NMR spectroscopic investigations confirmed the presence of type II beta-turns within each of the substituted peptides in contrast to the native sequence which contains a relatively low population of turn structure. In addition, an examination of singly substituted peptides suggests that an increase in the population of beta-turn structure within the amino-terminal Ser-Pro-Xaa-Xaa site also increased the formation of beta-turn structure in the carboxy-terminal (unmodified) Ser-Pro-Xaa-Xaa site; in comparison, substitution in the carboxy-terminal site did not influence structure in the remaining portion of the peptide. Overall, these results suggest that the structures formed could provide unique, preorganized linkers for the construction of novel DNA-interactive bisintercalators.
Subject(s)
RNA Polymerase II/chemistry , Circular Dichroism , Magnetic Resonance Spectroscopy , Models, Molecular , RNA Polymerase II/chemical synthesis , Repetitive Sequences, Nucleic AcidABSTRACT
The largest subunit of RNA polymerase II has a very interesting sequence in the C-terminus; that is, a tandem repeat sequence of Ser-Pro-Thr-Ser-Pro-Ser-Tyr consisted of proline residues and three kinds of residues having side-chain hydroxyl groups. Although lack of this tandem repeat is a lethal event in vivo, its functional role is unclear. The sequential polypeptide corresponding to this tandem repeat, poly(Ser-Pro-Thr-Ser-Pro-Ser-Tyr), was synthesized and its conformation was investigated by circular dichroism comparing to the monomeric heptapeptide. In addition, the theoretical conformational analysis based on the molecular mechanics was tried for the heptapeptide in the repeating unit and the periodic polyheptapeptide corresponding to the tandem repeat sequence. These results suggested the possibility that the tandem repeat contains a kind of super conformation composed of the repetitive turn structure in the native state. The characteristic repetitive turn structure would be the key of its function mechanism.
