Crystal structures reveal how distinct sites on the cysteine desulfurase IscS bind two different sulfur-acceptor proteins, IscU and TusA, to transfer sulfur atoms for iron-sulfur cluster biosynthesis and tRNA thiolation.
(2014) Modification of the wobble uridine in bacterial and mitochondrial tRNAs reading NNA/NNG triplets of 2-codon boxes, RNA Biology, 11:12, 1495-1507
Modifying RNA enzymes are highly specific for substrate-rRNA or tRNA-and the target position. In Escherichia coli, there are very few multisite acting enzymes, and only one rRNA/tRNA dual-specificity enzyme, pseudouridine synthase RluA, has been identified to date. Among the tRNA-modifying enzymes, the methyltransferase responsible for the m 2 A synthesis at purine 37 in a tRNA set still remains unknown. m 2 A is also present at position 2503 in the peptidyl transferase center of 23S RNA, where it is introduced by RlmN, a radical S-adenosyl-L-methionine (SAM) enzyme. Here, we show that E. coli RlmN is a dual-specificity enzyme that catalyzes methylation of both rRNA and tRNA. The DrlmN mutant lacks m 2 A in both RNA types, whereas the expression of recombinant RlmN from a plasmid introduced into this mutant restores tRNA modification. Moreover, RlmN performs m 2 A 37 synthesis in vitro using a tRNA chimera as a substrate. This chimera has also proved useful to characterize some tRNA identity determinants for RlmN and other tRNA modification enzymes. Our data suggest that RlmN works in a late step during tRNA maturation by recognizing a precise 3D structure of tRNA. RlmN inactivation increases the misreading of a UAG stop codon. Since loss of m 2 A 37 from tRNA is expected to produce a hyperaccurate phenotype, we believe that the error-prone phenotype exhibited by the DrlmN mutant is due to loss of m 2 A from 23S rRNA and, accordingly, that the m 2 A2503 modification plays a crucial role in the proofreading step occurring at the peptidyl transferase center.
The purpose of this study was to evaluate foot arch types of obese children and adolescents aged 9-16.5 years using both indirect and direct measures. Fifty-eight obese children/adolescents attending the paediatric endocrinology unit of the University Hospital "Lozano Blesa" in Zaragoza were selected as experimental subjects. Fifty-eight gender and age matched, normal-weight children/adolescents were selected as control subjects. To assess the medial longitudinal arch (MLA) height, which is used as a main reference for the diagnosis of flatfoot, footprints from both feet were collected (in both groups) and lateral weight-bearing radiographs of both feet were taken (of 49 of the 58 obese children). Footprint angle (FA) and the Chippaux-Smirak index (CSI) were calculated from the footprints. Talus-first metatarsal (TFMA) and calcaneal inclination angles (CIA) were obtained from lateral feet radiographs. In the normal-weight group, mean values of FA and CSI indicated a normal MLA. In the obese group, morphological flatfoot was identified. Comparison between both groups, by side and gender, showed a decrease of FA (p<0.001) and an increase of CSI (p<0.001) in obese subjects. Mean values of TFMA and CIA in the obese group indicated a lowering of the MLA. Obese children/adolescents between 9 and 16.5 years of age had significantly lower values of FA and higher CSI, related to a lower MLA. Radiographic parameters supported these findings and mean values were associated with a fall of this arch.
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