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Interaction of 42Sp50 with the vegetal RNA localization machinery in Xenopus laevis oocytes
Author(s) -
Loeber Jana,
Claußen Maike,
Jahn Olaf,
Pieler Tomas
Publication year - 2010
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2010.07878.x
Subject(s) - xenopus , microinjection , microbiology and biotechnology , biology , immunoprecipitation , oocyte , rna binding protein , rna , context (archaeology) , cytoplasm , messenger rna , untranslated region , embryo , genetics , gene , paleontology
Localization of a specific subset of maternal mRNAs to the vegetal cortex of Xenopus oocytes is important for the regulation of germ layer formation and germ cell development. It is driven by vegetal localization complexes that are formed with the corresponding signal sequences in the untranslated regions of the mRNAs and with a number of different so‐called localization proteins. In the context of the present study, we incorporated tagged variants of the known localization protein Vg1RBP into vegetal localization complexes by means of oocyte microinjection. Immunoprecipitation of the corresponding RNPs allowed for the identification of novel Vg1RBP‐associated proteins, such as the embryonic poly(A) binding protein, the Y‐box RNA‐packaging protein 2B and the oocyte‐specific version of the elongation factor 1α (42Sp50). Incorporation of 42Sp50 into localization RNPs could be confirmed by co‐immunoprecipitation of Vg1RBP and Staufen1 with myc‐tagged 42Sp50. Furthermore, myc‐42Sp50 was found to co‐sediment with the same two proteins in large, RNAse‐sensitive complexes, as well as to associate specifically with several vegetally localizing mRNAs but not with nonlocalized control RNAs. Finally, oocyte microinjection experiments reveal that 42Sp50 is a protein that shuttles between the nucleus and cytoplasm. Taken together, these observations provide evidence for a novel function of 42Sp50 in the context of vegetal mRNA transport in Xenopus oocytes. Structured digital abstract• MINT‐7994313 : epab (uniprotkb: Q98SP8 ) physically interacts ( MI:0915 ) with Vg1RBP (uniprotkb: O73932 ) by anti tag coimmunoprecipitation ( MI:0007 ) • MINT‐7994335 : 42Sp50 (uniprotkb: P17506 ) physically interacts ( MI:0915 ) with Vg1RBP (uniprotkb: O73932 ) by anti tag coimmunoprecipitation ( MI:0007 ) • MINT‐7994166 : Vg1RBP (uniprotkb: O73932 ) physically interacts ( MI:0914 ) with Vg1RBP (uniprotkb: O73932 ), 42Sp50 (uniprotkb: P17506 ), frgy2‐b (uniprotkb: P45441 ) and epab (uniprotkb: Q98SP8 ) by tandem affinity purification ( MI:0676 ) • MINT‐7994324 : frgy2‐b (uniprotkb: P45441 ) physically interacts ( MI:0915 ) with Vg1RBP (uniprotkb: O73932 ) by anti tag coimmunoprecipitation ( MI:0007 ) • MINT‐7994345 : 42Sp50 (uniprotkb: P17506 ) physically interacts ( MI:0914 ) with staufen (uniprotkb: Q5MNU4 ) and Vg1RBP (uniprotkb: O73932 ) by anti tag coimmunoprecipitation ( MI:0007 ) • MINT‐7994363 : 42Sp50 (uniprotkb: P17506 ), Vg1RBP (uniprotkb: O73932 ), staufen (uniprotkb: Q5MNU4 ) and 40LoVe (uniprotkb: Q6GM69 ) colocalize ( MI:0403 ) by cosedimentation through density gradient ( MI:0029 ) • MINT‐7994241 : Vg1RBP (uniprotkb: O73932 ) physically interacts ( MI:0914 ) with elrB (uniprotkb: Q91903 ), ElrA (uniprotkb: Q1JQ73 ), hnRNPI (uniprotkb: Q9PTS5 ), 40LoVe (uniprotkb: Q6GM69 ), staufen (uniprotkb: Q5MNU4 ) and Vg1RBP (uniprotkb: O73932 ) by tandem affinity purification ( MI:0676 )