Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS

Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS

Bibliographic Details
Main Author: Fu, Lihua
Language:English
Published: University of Cincinnati / OhioLINK 2015
Subjects:
Chemistry
tRNA
modification
Thermus thermophilus
HPLC
MS
xm5s2U
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537
id ndltd-OhioLink-oai-etd.ohiolink.edu-ucin1445342537
record_format oai_dc
spelling ndltd-OhioLink-oai-etd.ohiolink.edu-ucin14453425372021-08-03T06:33:29Z Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS Fu, Lihua Chemistry tRNA modification Thermus thermophilus HPLC MS xm5s2U Transfer RNAs (tRNAs) are essential adapter molecules for translation of the genetic code into a peptide sequence at the ribosome. The study of tRNA modifications are essential subjects of bioscience and are waiting to be explored in the areas of identification, function, structure, biosynthesis and the sequence of tRNA, and the locations of the post-transcriptional modification on tRNA sequences. <i>Thermus thermophilus </i> wild type and ΔTTHA 1897 were the model system that used to peruse a better understanding on tRNA modifications in this work. TTHA1897 is a putative gidA gene, which is one of the two crucial biosynthesis enzyme genes for xm<sup>5</sup>s<sup>2</sup>U in <i>E. coli</i>. ΔTTHA 1897 is mutant strain with disruption of tRNA gene TTHA1897 in <i>T. thermophilus</i>. Growth phenotype studies including growth rate and temperature sensitivity studies demonstrated that ΔTTHA1897 experienced a slower growth rate and non-survival in minimal medium. An HPLC-UV-MS study identified mnm<sup>5</sup>s<sup>2</sup>U in the wild type but not in ΔTTHA1897. Taking advantages of FT-ICRMS, the unique fragmentation pattern of xm<sup>5</sup>s<sup>2</sup>U was discovered with a proposed mechanism. An effective SRM method was developed and tested using <i>E. coli</i> tRNA nucleosides based on the unique fragmentation pattern of xm<sup>5</sup>s<sup>2</sup>U. A triple quadrupole mass spectrometer was also used to investigate a neutral loss method for the identification of xm<sup>5</sup>s<sup>2</sup>U nucleosides. 2015 English text University of Cincinnati / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537 http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Chemistry
tRNA
modification
Thermus thermophilus
HPLC
MS
xm5s2U
spellingShingle Chemistry
tRNA
modification
Thermus thermophilus
HPLC
MS
xm5s2U
Fu, Lihua
Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS
author Fu, Lihua
author_facet Fu, Lihua
author_sort Fu, Lihua
title Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS
title_short Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS
title_full Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS
title_fullStr Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS
title_full_unstemmed Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS
title_sort identification of trna modifications in t. thermophilus: wild type hb8 and mutant dttha1897 by lc-uv-ms/ms
publisher University of Cincinnati / OhioLINK
publishDate 2015
url http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537
work_keys_str_mv AT fulihua identificationoftrnamodificationsintthermophiluswildtypehb8andmutantdttha1897bylcuvmsms
_version_ 1719439144920285184

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