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Maturation of tRNA in Haloferax volcanii

Nist, Richard Neil
http://rave.ohiolink.edu/etdc/view?acc_num=osu1308066223

Abstract Details

2011, Doctor of Philosophy, Ohio State University, Microbiology.
Transfer RNA maturation in Archaea is a multi-faceted process requiring the activities of numerous enzymes in order to generate functional tRNAs from primary transcripts. Upon transcription, ribonuclease P cleaves the 5’ leader, tRNase Z cleaves the 3’ trailer, RNA-dependent and RNA-independent enzymes introduce modifications to bases and sugars, 3’ maturation is completed by the CCA-adding enzyme, and introns are removed. We examined the modification and intron removal stages of maturation for the Haloferax volcanii intron-containing pre-tRNA. H. volcanii tRNAMet possesses a 2’-O-methyl modification at position C34. A similar modification was characterized in pre-tRNATrp as being guided by a box C/D sRNP complex. Furthermore, just as sequence analysis and in vitro experiments have shown the intron-containing pre-tRNATrp to be a substrate for box C/D-guided modification, sequence analysis of intron-containing pre-tRNAMet suggests it is a substrate for box C/D-guided modification. We have determined not only that the C34 modification of tRNAMet is guided by a box C/D sRNP, but also that the intron-containing pre-tRNAMet appears to be a modification substrate in vivo. By the action of intron endonuclease, tRNA introns are removed in Archaea. This is a requisite step for the maturation of tRNATrp, tRNAMet, and tRNAGlnUUG in H. volcanii. Sequence analysis and previous mapping experiments suggest that there are additional substrates for this enzyme, including the 16S rRNA and 23S rRNA flanking sequences. Our data indicate that the 16S rRNA and 23S rRNA flanking sequences are cleaved by the intron endonuclease meaning there are substrates beyond the three tRNAs. Ribosomal protein L7ae has been at the forefront or background of each study conducted in this research. This K-turn stabilizing protein is involved in several facets of tRNA maturation including as a box C/D sRNP complex component required for 2’-O-methylation of tRNATrp and tRNAMet, as a component of archaeal RNase P, and as a box C/D sRNP complex component for the putative guide sRNA formed by the ligation of the 23S rRNA flanking sequences. Given the involvement of L7ae in these and other sRNP complexes, we attempted to capture L7ae-containing complexes from H. volcanii. We were able to demonstrate L7ae co-purification with each known and predicted class of L7ae-binding RNAs.
Charles Daniels (Advisor)
Juan Alfonzo (Committee Member)
Venkat Gopalan (Committee Member)
240 p.
Microbiology
intron endonuclease; sRNA; tRNA modification; archaea; box C/D

Recommended Citations

Citations

  • Nist, R. N. (2011). Maturation of tRNA in Haloferax volcanii [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1308066223

    APA Style (7th edition)

  • Nist, Richard. Maturation of tRNA in Haloferax volcanii. 2011. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1308066223.

    MLA Style (8th edition)

  • Nist, Richard. "Maturation of tRNA in Haloferax volcanii." Doctoral dissertation, Ohio State University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=osu1308066223

    Chicago Manual of Style (17th edition)

osu1308066223
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© 2011, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.