InterPro domain: IPR020094

Pseudouridine synthases catalyse the isomerisation of uridine to pseudouridine (Psi) in a variety of RNA molecules, and may function as RNA chaperones. Pseudouridine is the most abundant modified nucleotide found in all cellular RNAs. There are four distinct families of pseudouridine synthases that share no global sequence similarity, but which do share the same fold of their catalytic domain(s) and uracil-binding site and are descended from a common molecular ancestor. The catalytic domain consists of two subdomains, each of which has an alpha+beta structure that has some similarity to the ferredoxin-like fold (note: some pseudouridine synthases contain additional domains). The active site is the most conserved structural region of the superfamily and is located between the two homologous domains. These families are [ 1 , 2 ]:

• Pseudouridine synthase I, TruA.
• Pseudouridine synthase II, TruB, which contains and additional C-terminal PUA domain.
• Pseudouridine synthase RsuA. RluB, RluE and RluF are also part of this family.
• Pseudouridine synthase RluA. RluC and RluD belong to this family.
• Pseudouridine synthase TruD, which has a natural circular permutation in the catalytic domain, as well as an insertion of a family-specific alpha+beta subdomain.

TruA from Escherichia coli modifies positions uracil-38, U-39 and/or U-40 in tRNA [ 3 , 4 ]. TruA contains one atom of zinc essential for its native conformation and tRNA recognition and has a strictly conserved aspartic acid that is likely to be involved in catalysis [ 5 ]. These enzymes are dimeric proteins that contain two positively charged, RNA-binding clefts along their surface. Each cleft contains a highly conserved aspartic acid located at its centre. The structural domains have a topological similarity to those of other RNA-binding proteins, though the mode of interaction with tRNA appears to be unique.

This superfamily represents the Pseudouridine synthase I (TruA) N-terminal domain.

This entry also includes the N-terminal domain of several different pseudouridine synthases from family 3, including: RsuA (acts on small ribosomal subunit), RluB, RluE and RluF (act on large ribosomal subunit).

RsuA from Escherichia coli catalyses formation of pseudouridine at position 516 in 16S rRNA during assembly of the 30S ribosomal subunit [ 6 , 7 ]. RsuA consists of an N-terminal domain connected by an extended linker to the central and C-terminal domains. Uracil and UMP bind in a cleft between the central and C-terminal domains near the catalytic residue Asp 102. The N-terminal domain shows structural similarity to the ribosomal protein S4. Despite only 15% amino acid identity, the other two domains are structurally similar to those of the tRNA-specific psi-synthase TruA, including the position of the catalytic Asp. All four families of pseudouridine synthases share the same fold of their catalytic domain(s) and uracil-binding site.

RluB, RluE and RluF are homologous enzymes which each convert specific uridine bases in E. coli ribosomal 23S RNA to pseudouridine:

• RluB modifies uracil-2605.
• RluE modifies uracil-3457.
• RluF modifies uracil-2604 and to a lesser extent U-2605.

1. Role of cysteine residues in pseudouridine synthases of different families. Biochemistry 38, 13106-11
2. Enzymatic characterization and mutational studies of TruD--the fifth family of pseudouridine synthases. Arch. Biochem. Biophys. 489, 15-9
3. The structural basis for tRNA recognition and pseudouridine formation by pseudouridine synthase I. Nat. Struct. Biol. 7, 23-7
4. How U38, 39, and 40 of many tRNAs become the targets for pseudouridylation by TruA. Mol. Cell 26, 189-203
5. Transfer RNA-pseudouridine synthetase Pus1 of Saccharomyces cerevisiae contains one atom of zinc essential for its native conformation and tRNA recognition. Biochemistry 37, 7268-76
6. Structure of the 16S rRNA pseudouridine synthase RsuA bound to uracil and UMP. Nat. Struct. Biol. 9, 353-8
7. Structure of the pseudouridine synthase RsuA from Haemophilus influenzae. Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun. 61, 350-4