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Isolation and Properties of Escherichia coli 23S‐RNA Pseudouridine 1911, 1915, 1917 Synthase (RluD)
Author(s) -
Wrzesinski Jan,
Bakin Andrei,
Ofengand James,
Lane Byron G.
Publication year - 2000
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216540050176566
Subject(s) - pseudouridine , escherichia coli , ribosome , rna , gene , atp synthase , biology , biochemistry , 23s ribosomal rna , transfer rna , ribosomal rna , microbiology and biotechnology , enzyme , chemistry , genetics
All nine pseudouridine (psi) residues in Escherichia coli 23S RNA are in or very near the peptidyl transfer centre (PTC) of the ribosome. Five psi synthases catalyze synthesis of these nine psi's. Deletion of the gene for one psi synthase, RluD, which directs synthesis of three closely clustered psi's in the decoding site of the PTC, has a profound negative impact on cell growth. We describe the isolation, without amplification from a cloned coding element, of the triple-site modifying enzyme, RluD, the N-terminal sequence of which has been used to clone and express the corresponding gene, rluD. Unlike "expressed" RluD, which so far has not been shown to modify one (1911) of the three closely clustered sites (1911, 1915, 1917), "natural" RluD modifies all three sites; and unlike another pai synthase, RluA, natural RluD has greatly expanded modifying activity at low Mg concentrations. These properties of the expressed and natural forms of RluD are discussed.