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. 2018 Nov 28;8:17435. doi: 10.1038/s41598-018-35753-1

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© The Author(s) 2018

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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Dynamic interaction of PABP with ribosomes. (A–C) Fractions after sucrose-gradient centrifugation were analyzed by western blotting (WB: PABP, 40S and 60S) or northern blotting (NB: RNA). (A) PABP (60 pmol) alone (top panel) or a mixture of PABP (60 pmol) and the 40S subunit (30 pmol) (middle panel) or the 60S subunit (30 pmol) (bottom panel) was resolved by sucrose-gradient centrifugation. (B) A mixture of PABP (60 pmol), the poly(A) RNA (60 pmol), and the 40S subunit (30 pmol) or the 60S subunit (30 pmol) was resolved by sucrose-gradient centrifugation. (C) A mixture of PABP (60 pmol), the HA-Rluc-N RNA (60 pmol), and the 40S subunit (30 pmol) or the 60S subunit (30 pmol) was resolved by sucrose-gradient centrifugation. (D) Incubation with the poly(A) RNA (left panels) or HA-Rluc-N RNA (right panels) dissociated the 40S (upper panels) or 60S subunit (lower panels) from the PABP-His complex immobilized on the TALON resin (left three lanes, each panel). After incubation with RNA followed by washing, RNA/proteins were eluted with imidazole (500 mM) from the resin (right three lanes, each panel).