4JZS

Crystal structure of the Bacillus subtilis pyrophosphohydrolase BsRppH (E68A mutant)

4JZS の概要

• エントリーDOI: 10.2210/pdb4jzs/pdb
• 関連するPDBエントリー: 4JZT 4JZU 4JZV
• 分子名称: dGTP pyrophosphohydrolase, 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID (3 entities in total)
• 機能のキーワード: nudix hydrolase, rna pyrophosphohydrolase, rpph, cytosol, hydrolase
• 由来する生物種: Bacillus subtilis subsp. subtilis
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 41545.47

構造登録者

Piton, J.,Larue, V.,Thillier, Y.,Dorleans, A.,Pellegrini, O.,Li de la Sierra-Gallay, I.,Vasseur, J.J.,Debart, F.,Tisne, C.,Condon, C. (登録日: 2013-04-03, 公開日: 2013-05-08, 最終更新日: 2024-02-28)

主引用文献

Piton, J.,Larue, V.,Thillier, Y.,Dorleans, A.,Pellegrini, O.,Li de la Sierra-Gallay, I.,Vasseur, J.J.,Debart, F.,Tisne, C.,Condon, C.
Bacillus subtilis RNA deprotection enzyme RppH recognizes guanosine in the second position of its substrates.
Proc.Natl.Acad.Sci.USA, 110:8858-8863, 2013

PubMed Abstract: The initiation of mRNA degradation often requires deprotection of its 5' end. In eukaryotes, the 5'-methylguanosine (cap) structure is principally removed by the Nudix family decapping enzyme Dcp2, yielding a 5'-monophosphorylated RNA that is a substrate for 5' exoribonucleases. In bacteria, the 5'-triphosphate group of primary transcripts is also converted to a 5' monophosphate by a Nudix protein called RNA pyrophosphohydrolase (RppH), allowing access to both endo- and 5' exoribonucleases. Here we present the crystal structures of Bacillus subtilis RppH (BsRppH) bound to GTP and to a triphosphorylated dinucleotide RNA. In contrast to Bdellovibrio bacteriovorus RppH, which recognizes the first nucleotide of its RNA targets, the B. subtilis enzyme has a binding pocket that prefers guanosine residues in the second position of its substrates. The identification of sequence specificity for RppH in an internal position was a highly unexpected result. NMR chemical shift mapping in solution shows that at least three nucleotides are required for unambiguous binding of RNA. Biochemical assays of BsRppH on RNA substrates with single-base-mutation changes in the first four nucleotides confirm the importance of guanosine in position two for optimal enzyme activity. Our experiments highlight important structural and functional differences between BsRppH and the RNA deprotection enzymes of distantly related bacteria.

PubMed: 23610407
DOI: 10.1073/pnas.1221510110

実験手法

X-RAY DIFFRACTION (2.2 Å)