Full name: tRNA pseudouridine(32) synthase / Ribosomal large subunit pseudouridine synthase A
Synonym: YabO
GI: 76363883
Orf: yabO, b0058
COG: COG0564
UniProt: P0AA37
Structures: | 2I82 |
Complex:
Enzyme type: pseudouridine synthase
Position of modification - modification: t:32 - Y
l:746(746) - Y

Comments:

RluA forms pseudouridines at positions 746 in the loop of hairpin 35 of 23S ribosomal RNA (Domain II) and 32 in the loop of anticodon branch of a few E. coli tRNA. It reacts efficiently with full-length 23S RNA, a rRNA fragment encompassing the hairpin 35 (residues 1-847) and the mini tRNA comprising the anticodon stem-loop. RluA induces a dramatic reorganization of the RNA substrate allowing the target U to flip into the active site of the enzyme. Pseudouridylation occurs in single-stranded segments closed by a stem. The enzyme has no dependence on Mg2+, the rate of U-isomerisation is even faster in EDTA than in the presence of Mg2+. E. coli RluA belongs to the same subgroup of RNA pseudouridine synthases as E. coli, RluC, RluD and TruC as well as S. cerevisiae Pus5, Pus6, Pus8 and Pus9.

Protein sequence:

MGMENYNPPQEPWLVILYQDDHIMVVNKPSGLLSVPGRLEEHKDSVMTRIQRDYPQAESVHRLDMATSGV IVVALTKAAERELKRQFREREPKKQYVARVWGHPSPAEGLVDLPLICDWPNRPKQKVCYETGKPAQTEYE VVEYAADNTARVVLKPITGRSHQLRVHMLALGHPILGDRFYASPEARAMAPRLLLHAEMLTITHPAYGNS MTFKAPADF

Enzymatic activities:

Reaction Substrate Type Position
U:Y tRNA (t) Cys/GCA/prokaryotic cytosol 32
U:Y tRNA (t) Leu/UAA/prokaryotic cytosol 32
U:Y tRNA (t) Phe/GAA/prokaryotic cytosol 32
U:Y rRNA (r) LSU/23S/prokaryotic cytosol 746

Publications:

Title Authors Journal Details PubMed Id DOI
Mechanistic investigations of the pseudouridine synthase RluA using RNA containing 5-fluorouridine. Hamilton CS, Greco TM, Vizthum CA, Ginter JM, Johnston MV, Mueller EG Biochemistry [details] 17002302 -
A dual-specificity pseudouridine synthase: an Escherichia coli synthase purified and cloned on the basis of its specificity for psi 746 in 23S RNA is also specific for psi 32 in tRNA(phe). Wrzesinski J, Nurse K, Bakin A, Lane BG, Ofengand J RNA [details] 7493321 -
Crystal structure of pseudouridine synthase RluA: indirect sequence readout through protein-induced RNA structure. Hoang C, Chen J, Vizthum CA, Kandel JM, Hamilton CS, Mueller EG, Ferre-D'Amare AR Mol Cell [details] 17188032 -
Functional effect of deletion and mutation of the Escherichia coli ribosomal RNA and tRNA pseudouridine synthase RluA. Raychaudhuri S, Niu L, Conrad J, Lane BG, Ofengand J J Biol Chem [details] 10383384 -
Pre-steady-state kinetic analysis of the three Escherichia coli pseudouridine synthases TruB, TruA, and RluA reveals uniformly slow catalysis. Wright JR, Keffer-Wilkes LC, Dobing SR, Kothe U RNA [details] 21998096 -
The Handling of the Mechanistic Probe 5-Fluorouridine by the Pseudouridine Synthase TruA and Its Consistency with the Handling of the Same Probe by the Pseudouridine Synthases TruB and RluA. McDonald MK, Miracco EJ, Chen J, Xie Y, Mueller EG Biochemistry [details] 21142053 -

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