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

Fu, Lihua
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537

Abstract Details

2015, MS, University of Cincinnati, Arts and Sciences: Chemistry.
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. Thermus thermophilus 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 xm5s2U in E. coli. ΔTTHA 1897 is mutant strain with disruption of tRNA gene TTHA1897 in T. thermophilus. 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 mnm5s2U in the wild type but not in ΔTTHA1897. Taking advantages of FT-ICRMS, the unique fragmentation pattern of xm5s2U was discovered with a proposed mechanism. An effective SRM method was developed and tested using E. coli tRNA nucleosides based on the unique fragmentation pattern of xm5s2U. A triple quadrupole mass spectrometer was also used to investigate a neutral loss method for the identification of xm5s2U nucleosides.
Patrick Limbach, Ph.D. (Committee Chair)
William Heineman, Ph.D. (Committee Member)
Edward Merino, Ph.D. (Committee Member)
82 p.
Chemistry
tRNA; modification; Thermus thermophilus; HPLC; MS; xm5s2U

Recommended Citations

Citations

  • Fu, L. (2015). Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS [Master's thesis, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537

    APA Style (7th edition)

  • Fu, Lihua. Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS. 2015. University of Cincinnati, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537.

    MLA Style (8th edition)

  • Fu, Lihua. "Identification of tRNA modifications in T. thermophilus: wild type HB8 and mutant DTTHA1897 by LC-UV-MS/MS." Master's thesis, University of Cincinnati, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1445342537

    Chicago Manual of Style (17th edition)

ucin1445342537
370
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