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. 2012 Oct 1;23(19):3863–3872. doi: 10.1091/mbc.E12-02-0142

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© 2012 Brenna et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — Identification of a WC-1 LXXLL motif involved in the association with HAT. (A) Schematic representation of a generic nuclear receptor (NR) and WC-1 structural organization, evidencing architectural and functional similarities. (B) Multialignment of the regions containing the LXXLL motifs within Neurospora WC-1 and several WC-1–like proteins present in the National Center for Biotechnology Information databank (LLDLL and LAQLL, indicated by a dashed line). Consensus analysis reveals conservation only for the LXXLL motif present at position 1008–1012 in Neurospora WC-1. Organisms and protein accession numbers: Cryptococcus neoformans, XP_567995.1; Cryptococcus gattii, XP_003193431.1; Neurospora crassa, CAA63964; Sordaria macrospora, XP_003351224.1; Podospora anserina, CAD60767; Verticillium albo-atrum, XP_003001986.1; Colletotrichum higginsianum, CCF31796.1; Glomerella graminicola, EFQ30482.1; Nectria haematococca, XP_003040715.1; Fusarium oxysporum, ABY47609; Gibberella fujikuroi, CAO85915; Gibberella zae, XP_388117; Hypocrea jecorina, AAV80185; Trichoderma atroviride, AAU14171.1; Metarhizium anisopliae, EFY99524.1; Metarhizium acridum, EFY92414; Magnaporthe oryzae, XP_360995.2; Thielavia terrestris, XP_003654033; Chaetomium globosum XP_001219613; Myceliophthora thermophila, XP_003665493.1; Botryotinia fuckeliana, XP_001547999.1; Sclerotinia sclerotiorum, XP_001586924.1; Cercospora zeae-maydis, AEH41590.1; Mycosphaerella graminicola, EGP83257.1; Phaeosphaeria avenaria, ACS74812.1; Cochliobolus miyabeanus, BAF35570.1; Leptosphaeria maculans, CBX98033.1; Pyrenophora tritici-repentis, XP_001933567.1; Tuber borchii, CAE01390.1; M., Tuber melanosporum, CAZ83744.1; Aspergillus niger, XP_001395039.1; Aspergillus nidulans, XP_661040.1. (C) Schematic representation of the domains in the two WC-1 mini versions and WT and point mutation mutant (Mut LXXAA) used in the pull-down experiments. The LAQLL mutagenized version is changed in LAQAA. (D) Recombinant proteins (C) were used in pull-down assays to evaluate their interaction with a radiolabeled Sc-Gcn5p. A representative Coomassie-stained gel and its autoradiography are shown (top). Quantification of the relative amount of 35S-ScGcn5p retained by the two recombinant products. The signal corresponding to WC-1 mini/ScGcn5p interaction was set to 100% (bottom). The data are the mean ± SEM from three independent experiments.