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. 2020 Jan 8;11:122. doi: 10.1038/s41467-019-13991-9

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Schematic model of mRNA1 before and after the endonucleolytic cleavages producing B4 RNAs. 5′-NGD resulting RNAs are shown here covered by two ribosomes and processed by Xrn1 to 48-nt RNAs (see also Supplementary Fig. 4 for 5′-NGD RNAs covered by three ribosomes). b 8% PAGE followed by northern blot analysis using probe prG showing steady-state levels of RNAs in the indicated mutant strains (left panel). Same membrane has been probed with prA as a ladder (right panel), and sizes of B5 (77nt), B4 (71 nt) and B1 (47nt) are indicated. The 5S rRNA served as a loading control. c Primer extension experiments using probe prG to determine the 5’-end of RNAs (see also Supplementary Fig. 5). d Schematic model of ribosome positioning on mRNA1 before and after the unique endonucleolytic cleavage producing B4 RNAs, localized 8 nts upstream of the first P-site nt. The position of disomes on the resulting 48-nt 5’-NGD RNA is shown with the distal ribosome having 1 nt in the A site (see also Supplementary Fig. 4). Source data are provided as a Source Data file.