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Post-Transcriptional Regulation of Selenoprotein S

Cockman, Eric Michael
http://orcid.org/0000-0002-9415-7636
http://rave.ohiolink.edu/etdc/view?acc_num=case1562593531805034

Abstract Details

2019, Doctor of Philosophy, Case Western Reserve University, Molecular Medicine.
Selenoproteins are a unique class of proteins that contain the 21st amino acid, selenocysteine (Sec). Addition of Sec into a protein is achieved by recoding of the UGA stop codon. All 25 mammalian selenoprotein mRNAs possess a 3’ UTR stem-loop structure, the Selenocysteine Insertion Sequence (SECIS), which is required for Sec incorporation. It is widely believed that the SECIS is the major RNA element that controls Sec insertion, however recent findings in our lab suggest otherwise for Selenoprotein S (SelS). Here we report that the first 60 nucleotides of the SelS 3’ UTR contains a proximal stem loop (PSL) and a conserved sequence we have named the SelS Positive UGA Recoding (SPUR) element. We developed a SelS-V5/UGA surrogate assay for UGA recoding, which was validated by mass spectrometry to be an accurate measure of Sec incorporation in cells. Using this assay, we show that point mutations in the SPUR greatly reduce recoding in the reporter; thus, the SPUR is required for readthrough of the UGA-Sec codon. In contrast, deletion of the PSL increased Sec incorporation. This effect was reversed when the PSL was replaced with other stem-loops or a linear sequence, suggesting that the PSL does not play an active role in Sec insertion. Additional studies revealed that the position of the SPUR relative to the UGA-Sec codon is important for optimal UGA recoding. Our identification of the SPUR element in the SelS 3’ UTR reveals a more complex regulation of Sec incorporation than previously realized.
Donna Driscoll, Ph.D. (Advisor)
125 p.
Biology; Cellular Biology; Molecular Biology
selenoproteins; selenoprotein S; selenocysteine; mRNA translation; 3 untranslated region

Recommended Citations

Citations

  • Cockman, E. M. (2019). Post-Transcriptional Regulation of Selenoprotein S [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1562593531805034

    APA Style (7th edition)

  • Cockman, Eric. Post-Transcriptional Regulation of Selenoprotein S. 2019. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1562593531805034.

    MLA Style (8th edition)

  • Cockman, Eric. "Post-Transcriptional Regulation of Selenoprotein S." Doctoral dissertation, Case Western Reserve University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=case1562593531805034

    Chicago Manual of Style (17th edition)

case1562593531805034
289
© 2019, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.