3DH3

Crystal Structure of RluF in complex with a 22 nucleotide RNA substrate

Summary for 3DH3

• Entry DOI: 10.2210/pdb3dh3/pdb
• Descriptor: Ribosomal large subunit pseudouridine synthase F, stem loop fragment of E. Coli 23S RNA (3 entities in total)
• Functional Keywords: protein-rna complex, s4 domain, alpha/beta protein, isomerase, rna-binding, rrna processing, isomerase-rna complex, isomerase/rna
• Biological source: Escherichia coli
• Total number of polymer chains: 8
• Total formula weight: 158831.98

Authors

Alian, A.,DeGiovanni, A.,Stroud, R.M.,Finer-Moore, J.S. (deposition date: 2008-06-16, release date: 2009-04-28, Last modification date: 2023-08-30)

Primary citation

Alian, A.,DeGiovanni, A.,Griner, S.L.,Finer-Moore, J.S.,Stroud, R.M.
Crystal structure of an RluF-RNA complex: a base-pair rearrangement is the key to selectivity of RluF for U2604 of the ribosome.
J.Mol.Biol., 388:785-800, 2009

PubMed Abstract: Escherichia coli pseudouridine synthase RluF is dedicated to modifying U2604 in a stem-loop of 23S RNA, while a homologue, RluB, modifies the adjacent base, U2605. Both uridines are in the same RNA stem, separated by approximately 4 A. The 3.0 A X-ray crystal structure of RluF bound to the isolated stem-loop, in which U2604 is substituted by 5-fluorouridine to prevent catalytic turnover, shows RluF distinguishes closely spaced bases in similar environments by a selectivity mechanism based on a frameshift in base pairing. The RNA stem-loop is bound to a conserved binding groove in the catalytic domain. A base from a bulge in the stem, A2602, has folded into the stem, forcing one strand of the RNA stem to translate by one position and thus positioning U2604 to flip into the active site. RluF does not modify U2604 in mutant stem-loops that lack the A2602 bulge and shows dramatically higher activity for a stem-loop with a mutation designed to facilitate A2602 refolding into the stem with concomitant RNA strand translation. Residues whose side chains contact rearranged bases in the bound stem-loop, while conserved among RluFs, are not conserved between RluFs and RluBs, suggesting that RluB does not bind to the rearranged stem loop.

PubMed: 19298824
DOI: 10.1016/j.jmb.2009.03.029

Experimental method

X-RAY DIFFRACTION (3 Å)