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. Author manuscript; available in PMC: 2014 Jul 2.
Published in final edited form as: Nature. 2013 Nov 17;505(7481):112–116. doi: 10.1038/nature12731

Figure 4. Nug2 release from the pre-60S subunit is linked to Nmd3 recruitment.

Figure 4

(a) Affinity-purification of the indicated TAP-tagged pre-60S factors from NUG2 or nug2K328R mutant cells. (*) position of Rei1 identified by mass spectrometry. (b) Affinity-purification of Arx1-TAP from NUG2, nug2K328R and nug2G369A cells. (c) Affinity-purification of Rix1-TAP from sAid-Nug2-sAid degron strain after time-dependent auxin treatment. (d) Affinity-purification of Nug2-TAP particles with (lane 1) or without (lane 2) subsequent Rix1-Flag immunodepletion. (a-d) SDS-PAGE and Coomassie staining (upper panel) and Western blotting using the indicated antibodies (lower panel). Protein bands indicated were identified by mass spectrometry. All affinity-purifications were performed at least twice, yielding highly reproducible datasets. (e) Model of pre-60S subunit maturation starting from the Rix1-particle with final Nmd3-Crm1-RanGTP recruitment.