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Study of the L13a residues required for ribosomal function

Das, Priyanka
http://rave.ohiolink.edu/etdc/view?acc_num=csu1331762160

Abstract Details

2012, Doctor of Philosophy in Regulatory Biology, Cleveland State University, College of Sciences and Health Professions.
Ribosome biogenesis, a fundamental process, occurs in the nucleolus. It involves incorporation and association of ribosomal proteins (r-proteins) in the ribosomal subunit. Despite its obligatory and critical role in cellular function, the explicit mechanism of incorporation of different r-proteins into the pre-ribosome is not well understood. Using mammalian cell and r-protein L13a as our model, this study addresses the function and mechanism of r-protein incorporation during ribosome maturation. Published results from our laboratory showed the requirement of the release of L13a from the 60S ribosome to silence a cohort of inflammatory proteins directly at the level of translation thus showing the significant potential of this mechanism to resolve inflammation. To get further insight into the mechanism of its release, it is essential to identify the domain of L13a and the subcellular site required for ribosome incorporation. Homology modeling of human L13a with the crystal structure of prokaryotic L13, predicted some amino acid residues that could bind to ribosomal RNA (rRNA). Consistent with this model, a combined experimental approach involving ribosome incorporation assay of recombinant L13a and RNA immunoprecipitation have identified Arg at position 68 and Arg-Lys-Arg at position 59-60-61 as potential interaction site with 60S subunit. We have performed immunofluorescence studies to test whether the incorporation defective mutant L13a failed to translocate to the nucleolus, the site of ribosome biogenesis. L13a harboring the mutation of Arg at position 68 to Ala translocate to the nucleolus, but however alters the nucleolar morphology. In contrast L13a with the mutation of Arg-Lys-Arg at position 59-60-61 to Ala-Ala-Ala is nucleolar translocation incompetent. These studies also identified that incorporation of L13a during ribosome biogenesis occurs at the stage of 90S pre-ribosome formation. Previous studies from our laboratory showed an essential role of L13a in rRNA methylation. In these studies we identified L13a as an interacting partner of Fibrillarin containing C/D box snoRNP complex, thus showing a strong rationale of L13a as a rRNA methylation factor. Most importantly, these studies also showed that incorporation of L13a in the 60S subunit is restricted only during ribosome biogenesis and not after maturation.
Barsanjit Mazumder, Ph.D (Advisor)
Crystal Weyman, Ph.D (Committee Member)
Anton Komar, Ph.D (Committee Member)
Jaharul Haque, Ph.D. (Committee Member)
Girish Shukla, Ph.D. (Committee Member)
Amin Zhou, Ph.D. (Committee Member)
Molecular Biology
L13a; ribosome biogenesis; rRNA methylation;

Recommended Citations

Citations

  • Das, P. (2012). Study of the L13a residues required for ribosomal function [Doctoral dissertation, Cleveland State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=csu1331762160

    APA Style (7th edition)

  • Das, Priyanka. Study of the L13a residues required for ribosomal function. 2012. Cleveland State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=csu1331762160.

    MLA Style (8th edition)

  • Das, Priyanka. "Study of the L13a residues required for ribosomal function." Doctoral dissertation, Cleveland State University, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=csu1331762160

    Chicago Manual of Style (17th edition)

csu1331762160
895
© 2012, all rights reserved.
This open access ETD is published by Cleveland State University and OhioLINK.