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. 2017 Jun 9;7:3169. doi: 10.1038/s41598-017-03034-y

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© The Author(s) 2017

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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Association of Utp21p with pre-rRNAs under the expression of different PWP2 alleles. (A) Schematic representation of the rRNA processing pathway during maturation of the 40S ribosomal subunit in yeast. Probes used in northern blot analysis are depicted in red. The rRNAs that were detected in our assays are labelled in red. Alternative options for an initial cleavage of the 35S at positions A0, A1, A2, or A3 is shown. All possible cleavage options during the maturation of the 40S particle are depicted in the A2 and A3 pathways. (B) The yeast strain yCMS3-1a harbouring empty plasmid, wild-type PWP2 or truncated forms of the gene PWP2, were cultivated in SCD-U medium. Twelve percent from the WCL and 16.5% from the Utp21p-TAP-purified fraction were analysed (with the exception of the yMJH1-1a, where 25% of the WCL was analysed). The different rRNA species were resolved on either agarose gels (top three panels) or urea-acrylamide gels (two panels at the bottom) and analysed by northern blotting using the oligonucleotides: #2921 (upper panel), #207 (second panel from the top), #3839 (third panel from the top), #3468 (four panel from the top) or #3470 (lower panel). The hybridisation sites of the different probes are predicted in panel A. The apparent size of the detected rRNAs is indicated at the right side. (C) RNAs co-purified with Utp21p-TAP were compared with the total RNAs present in cell lysates.