RRC ID 27711
Author Ishida K, Kunibayashi T, Tomikawa C, Ochi A, Kanai T, Hirata A, Iwashita C, Hori H.
Title Pseudouridine at position 55 in tRNA controls the contents of other modified nucleotides for low-temperature adaptation in the extreme-thermophilic eubacterium Thermus thermophilus.
Journal Nucleic Acids Res.
Abstract Pseudouridine at position 55 (Ψ55) in eubacterial tRNA is produced by TruB. To clarify the role of the Ψ55 modification, we constructed a truB gene disruptant (ΔtruB) strain of Thermus thermophilus which is an extreme-thermophilic eubacterium. Unexpectedly, the ΔtruB strain exhibited severe growth retardation at 50 °C. We assumed that these phenomena might be caused by lack of RNA chaperone activity of TruB, which was previously hypothetically proposed by others. To confirm this idea, we replaced the truB gene in the genome with mutant genes, which express TruB proteins with very weak or no enzymatic activity. However the growth retardation at 50 °C was not rescued by these mutant proteins. Nucleoside analysis revealed that Gm18, m(5)s(2)U54 and m(1)A58 in tRNA from the ΔtruB strain were abnormally increased. An in vitro assay using purified tRNA modification enzymes demonstrated that the Ψ55 modification has a negative effect on Gm18 formation by TrmH. These experimental results show that the Ψ55 modification is required for low-temperature adaptation to control other modified. (35)S-Met incorporation analysis showed that the protein synthesis activity of the ΔtruB strain was inferior to that of the wild-type strain and that the cold-shock proteins were absence in the ΔtruB cells at 50°C.
Volume 39(6)
Pages 2304-18
Published 2011-3
DOI 10.1093/nar/gkq1180
PII gkq1180
PMID 21097467
PMC PMC3064792
MeSH Adaptation, Physiological Bacterial Proteins / genetics Bacterial Proteins / metabolism* Intramolecular Transferases / genetics Intramolecular Transferases / metabolism* Methionine / metabolism Molecular Chaperones / metabolism Mutation Nucleotides / chemistry Nucleotides / metabolism Pseudouridine / metabolism* RNA Processing, Post-Transcriptional RNA, Transfer / chemistry RNA, Transfer / metabolism* RNA, Transfer, Met / chemistry Temperature* Thermus thermophilus / enzymology* Thermus thermophilus / genetics Thermus thermophilus / growth & development tRNA Methyltransferases / metabolism
IF 11.561
Times Cited 23
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
DNA material pUC18-HTK (with promoter) (RDB03436)