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. 2012 Apr 13;31(11):2579–2589. doi: 10.1038/emboj.2012.85

Figure 2.

Figure 2

Nonfunctional ribosomes are selectively ubiquitinated and degraded in an Mms1-dependent manner. (A) Northern blotting of 18-nt- or MS2-tagged 25S rRNAs. The total RNAs were isolated from cells carrying the indicated plasmids after transcriptional shut-off. These RNAs were cleaved with RNase H (Supplementary Figure S7A). Northern hybridization was performed using a probe that detected both tagged RNAs. (B, C) Immunoblotting of ribosomes purified from cells expressing the indicated tagged rRNAs and Myc–ubiquitin. (B) Ribosomes were Rpl28–Flag immunopurified. In lane 6, an empty vector was used instead of pMyc–Ubi. In lane 7, a wild-type strain with untagged Rpl28 was used. (C) GST–MS2 affinity purification of the ribosomal fraction sedimented by sucrose density gradient centrifugation. In lane 6, an 18-nt-tagged wild-type rRNA was expressed as the negative control. Figure source data can be found with the Supplementary data.