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Formation of a Fuss/Med protein complex and its translocation into the nucleus.

By Susanne Fischer (312335), Florian Bayersdorfer (312337), Eva Harant (312338), Renate Reng (312339), Stephanie Arndt (312340), Anja-Katrin Bosserhoff (312341) and Stephan Schneuwly (312342)

Abstract

<p>(A) Yeast Two Hybrid experiment showing physical interaction of Fuss and Med: <i>pPC97-Fos + pPC86-Jun</i>: interaction control; <i>pCL1+ pPC86</i>: Gal4- growth control and empty prey vector; <i>pdbLeu-Fuss + pPC86</i>: negative control; <i>pdbLeu-Fuss + pPC86-Med</i>: positive interaction of Fuss and Med. (B) Coimmunoprecipitations: in lysates from S2-cells co-expressing HA-Fuss together with FLAG-Med or FLAG-Mad, HA-Fuss co-precipitates with FLAG-Med but not with FLAG-Mad. In lysates from S2-cells co-expressing FLAG-Fuss with HA-Med or HA-Mad, HA-Med co-precipitates with FLAG-Fuss and HA-Mad does not. (C) Wing of a male fly of the genotype <i>A9-Gal4; UAS-fuss-GFP</i>. Overexpression of <i>fuss-GFP</i> leads to truncations of L2 and L5 veins and a loss of the p-cv. (D–E”) Upon co-expression of Med, Fuss-GFP is partially translocated into the nucleus while the cytoplasmic fraction of the protein is reduced. Confocal scans of L3 wing discs stained with anti-GFP (D, E) and anti-Histone (D’, E’) antibodies. (D–D”) <i>A9-Gal4; UAS-fuss-GFP</i>. The Fuss-GFP fusion protein is mainly localised in the cytoplasm. (E) <i>A9-Gal4; UAS-fuss-GFP; UAS-Med</i>. Arrows emphasize some of the cells that clearly show nuclear localisation of the Fuss-GFP fusion protein. (F–G”) Fuss does not inhibit the nuclear translocation of pMad. Confocal scans of L3 wing discs stained with anti-phospho-SMAD1/5 (F, G) and anti-Histone (F’, G’) antibodies. (F–F”) <i>A9-Gal4.</i> (G–G”) <i>A9-Gal4; UAS-fussB</i>. All Scale bars: 10 µm.</p

Topics: Developmental Biology, translocation
Year: 2013
DOI identifier: 10.1371/journal.pone.0042349.g005
OAI identifier: oai:figshare.com:article/266287
Provided by: FigShare
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