| RRC ID |
5823
|
| Author |
Tomikawa C, Yokogawa T, Kanai T, Hori H.
|
| Title |
N7-Methylguanine at position 46 (m7G46) in tRNA from Thermus thermophilus is required for cell viability at high temperatures through a tRNA modification network.
|
| Journal |
Nucleic Acids Res
|
| Abstract |
N(7)-methylguanine at position 46 (m(7)G46) in tRNA is produced by tRNA (m(7)G46) methyltransferase (TrmB). To clarify the role of this modification, we made a trmB gene disruptant (DeltatrmB) of Thermus thermophilus, an extreme thermophilic eubacterium. The absence of TrmB activity in cell extract from the DeltatrmB strain and the lack of the m(7)G46 modification in tRNA(Phe) were confirmed by enzyme assay, nucleoside analysis and RNA sequencing. When the DeltatrmB strain was cultured at high temperatures, several modified nucleotides in tRNA were hypo-modified in addition to the lack of the m(7)G46 modification. Assays with tRNA modification enzymes revealed hypo-modifications of Gm18 and m(1)G37, suggesting that the m(7)G46 positively affects their formations. Although the lack of the m(7)G46 modification and the hypo-modifications do not affect the Phe charging activity of tRNA(Phe), they cause a decrease in melting temperature of class I tRNA and degradation of tRNA(Phe) and tRNA(Ile). (35)S-Met incorporation into proteins revealed that protein synthesis in DeltatrmB cells is depressed above 70 degrees C. At 80 degrees C, the DeltatrmB strain exhibits a severe growth defect. Thus, the m(7)G46 modification is required for cell viability at high temperatures via a tRNA modification network, in which the m(7)G46 modification supports introduction of other modifications.
|
| Volume |
38(3)
|
| Pages |
942-57
|
| Published |
2010-1-1
|
| DOI |
10.1093/nar/gkp1059
|
| PII |
gkp1059
|
| PMID |
19934251
|
| PMC |
PMC2817472
|
| MeSH |
Aminoacylation
Bacterial Proteins / biosynthesis
Bacterial Proteins / chemistry
Gene Deletion
Guanine / analogs & derivatives*
Guanine / metabolism
Hot Temperature*
Methionine / metabolism
Nucleic Acid Denaturation
RNA Processing, Post-Transcriptional
RNA, Transfer / chemistry*
RNA, Transfer / metabolism
RNA, Transfer, Phe / chemistry
RNA, Transfer, Phe / metabolism
Thermus thermophilus / enzymology*
Thermus thermophilus / genetics
Thermus thermophilus / growth & development
tRNA Methyltransferases / genetics
tRNA Methyltransferases / metabolism*
|
| IF |
11.502
|
| Times Cited |
59
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| DNA material |
Thermus thermophilus HB8 gene disruption plasmid TDs08C11 (THR023259) |
