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A second function for pseudouridine synthases: A point mutant of RluD unable to form pseudouridines 1911, 1915 and 1917 in Escherichia coli 23S ribosomal RNA restores normal growth to an RluD-minus strain

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title A second function for pseudouridine synthases: A point mutant of RluD unable to form pseudouridines 1911, 1915 and 1917 in Escherichia coli 23S ribosomal RNA restores normal growth to an RluD-minus strain
 
Creator Gutgsell, N.S
Campo, M.D.
Chaudhuri, Saumya Ray
Ofengand, J.
 
Subject QR Microbiology
 
Description This laboratory previously showed that truncation of the gene for RluD, the Escherichia coli pseudouridine synthase responsible for synthesis of 23S rRNA pseudouridines 1911, 1915, and 1917, blocks pseudouridine formation and inhibits growth. We now show that RluD mutants at the essential aspartate 139 allow these two functions of RluD to be separated. In vitro, RluD with aspartate 139 replaced by threonine or asparagine is completely inactive. In vivo, the growth defect could be completely restored by transformation of an RluD-inactive strain with plasmids carrying genes for RluD with aspartate 139 replaced by threonine or asparagine. Pseudouridine sequencing of the 23S rRNA from these transformed strains demonstrated the lack of these pseudouridines. Pseudoreversion, which has previously been shown to restore growth without pseudouridine formation by mutation at a distant position on the chromosome, was not responsible because transformation with empty vector under identical conditions did not alter the growth rate.
 
Publisher Cambridge University Press/ RNA Society
 
Date 2011-07
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://crdd.osdd.net/open/861/1/saumya2001.pdf
Gutgsell, N.S and Campo, M.D. and Chaudhuri, Saumya Ray and Ofengand, J. (2011) A second function for pseudouridine synthases: A point mutant of RluD unable to form pseudouridines 1911, 1915 and 1917 in Escherichia coli 23S ribosomal RNA restores normal growth to an RluD-minus strain. RNA., 7 (7). pp. 990-998.
 
Relation http://crdd.osdd.net/open/861/