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Isolation and Properties of Escherichia coli 23S‐RNA Pseudouridine 1911, 1915, 1917 Synthase (RluD)

Isolation and Properties of Escherichia coli 23S-RNA Pseudouridine 1911, 1915, 1917 Synthase (RluD) Jan Wrzesinski,1 Andrei Bakin,2 James Ofengand,3 and Byron G. Lane4 Polish Academy of Sciences, Institute of Bioorganic Chemistry, ul. Noskowskiego 12/14, 61-704, Poznan, Poland 2 Division of Hematology-Oncology, Vanderbilt University, 22nd Avenue South, Nashville, Tennessee 37232-5536 3 University of Miami, Department of Biochemistry and Molecular Biology, School of Medicine, Miami, Florida 33101 4 University of Toronto, Department of Biochemistry, Toronto, Ontario, Canada M5S 1A8 Summary All nine pseudouridine (W ) residues in Escherichia coli 23S RNA are in or very near the peptidyl transfer centre (PTC) of the ribosome. Five W synthases catalyze synthesis of these nine W ’s. Deletion of the gene for one W synthase, RluD, which directs synthesis of three closely clustered W ’s in the decoding site of the PTC, has a profound negative impact on cell growth. We describe the isolation, without ampli cation from a cloned coding element, of the triplesite 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 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png IUBMB Life Wiley

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