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oai:figshare.com:article/34047638 2026-10-01T17:26:13Z category_1 category_4 category_8 category_13 category_14 category_873 category_734 category_61 category_64 category_134 portal_5 item_type_1 month_year_10_2026

<p>The main stem loop of the inhibitory structure of <i>RPL22B</i>i was necessary but not sufficient for splicing inhibition.</p> Kateřina Abrhámová (4739808) Alexandra Gredová (25157630) Karolína Navrátilová (25157633) Mohamed Boumaiza (5636147) Petr Folk (209900) Biophysics Biochemistry Microbiology Genetics Molecular Biology Chemical Sciences not elsewhere classified Biological Sciences not elsewhere classified Developmental Biology Cancer Virology intergenic regulation mediated first catalytic step 5 ’ ss div >< p additional stem loops tested splicing efficiency saccharomyces cerevisiae </ rather stalled splicing enhanced splicing inhibition intron binding using predicted stem alternate stem p structure permitted splicing b </ structural ensemble rpl22 </ ribosomal roles ribosomal proteins regulatory mechanism prevent recognition predicted features particular type others described mutagenesis identified mrna behaves later stage hybrid system fulfill non containing reporter branch point binding capacity allosteric switch <p>The experiment was done and the figure was formatted as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0359197#pone.0359197.g002" target="_blank">Fig 2</a>. (A) The inhibitory mechanism required that specific structural details of the I structure remain intact. In the mutants analyzed in this set, the main stem loop is predicted to form. Melting of Loop2 of the I structure (BPmut4; see panel (C)) renders the structure splicing permissive, while ablating the loop (dLoop2) retains the Rpl22-dependent inhibitory potential. Although the stability of the main stem loop should be lowered in Mutclamp2, it is apparently still sufficient to keep the I structure Rpl22 responsive. (B) Summary of the reporter experiments in (A) and types of stem loop arrangement predicted by RNAshapes. pre-mRNA accumulation was approximated on a scale of ‘+++’ to ‘-.’ The secondary structures were predicted using RNAshapes [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0359197#pone.0359197.ref054" target="_blank">54</a>]. The structures were sorted according to the stem loop arrangement involving the conserved region and its complementary elements (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0359197#pone.0359197.g001" target="_blank">Fig 1</a>), and the number of structure types classified as I, P, or O (other) was expressed in %. See text and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0359197#pone.0359197.s001" target="_blank">S1</a>-<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0359197#pone.0359197.s003" target="_blank">S3 Figs</a> for additional information. (C) Detail of the predicted secondary structure of <i>RPL22B</i>i in the I conformation. The regions referred to in the text are indicated (in italics) together with the mutants (in plain text). (D) The main stem loop was not sufficient to convey splicing inhibition in an unrelated intron. <i>The RPS17B</i> intron without or with the main stem loop inserted was tested as in (A). M indicates the m.w. marker lane.</p> 2026-10-01T17:25:45Z Image Figure 10.1371/journal.pone.0359197.g004 https://figshare.com/articles/figure/_p_The_main_stem_loop_of_the_inhibitory_structure_of_i_RPL22B_i_i_was_necessary_but_not_sufficient_for_splicing_inhibition_p_/34047638 CC BY 4.0