1XST

Solution structure of E.coli RNase P RNA P4 stem, U69A mutation, complexed with cobalt (III) hexammine.

Summary for 1XST

• Entry DOI: 10.2210/pdb1xst/pdb
• Related: 1F6X 1F78 1XSU
• Descriptor: RNA (27-MER), COBALT HEXAMMINE(III) (2 entities in total)
• Functional Keywords: ribonuclease p rna, ribozyme, transfer rna processing, p4 stem, u69a mutant, metal binding site, metal complex, cobalt (iii) hexammine complex, rna
• Total number of polymer chains: 1
• Total formula weight: 8817.30

Authors

Schmitz, M. (deposition date: 2004-10-20, release date: 2004-12-28, Last modification date: 2024-05-29)

Primary citation

Schmitz, M.
Change of RNase P RNA function by single base mutation correlates with perturbation of metal ion binding in P4 as determined by NMR spectroscopy
Nucleic Acids Res., 32:6358-6366, 2004

PubMed Abstract: The solution structures of two 27 nt RNA hairpins and their complexes with cobalt(III)-hexammine [Co(NH(3))(6)(3+)] were determined by NMR spectroscopy. The RNA hairpins are variants of the P4 region from Escherichia coli RNase P RNA: a U-to-A mutant changing the identity of the bulged nucleotide, and a U-to-C, C-to-U double mutant changing only the bulge position. Structures calculated from NMR constraints show that the RNA hairpins adopt different conformations. In the U-to-C, C-to-U double mutant, the conserved bulged uridine in the P4 wild-type stem is found to be shifted in the 3'-direction by one nucleotide when compared with the wild-type structure. Co(NH(3))(6)(3+) is used as a spectroscopic probe for Mg(H(2)O)(6)(2+) binding sites because both complexes have octahedral symmetry and have similar radii. Intermolecular NOE crosspeaks between Co(NH(3))(6)(3+) and RNA protons were used to locate the site of Co(NH(3))(6)(3+) binding to both RNA hairpins. The metal ion binds in the major groove near a bulge loop in both mutants, but is shifted 3' by about one base pair in the double mutant. The change of the metal ion binding site is compared with results obtained on corresponding mutant RNase P RNA molecules as reported by Harris and co-workers (RNA, 1, 210-218).

PubMed: 15576680
DOI: 10.1093/nar/gkh961

Experimental method

SOLUTION NMR