caa a22 4500 397573472 CHVBK 20180308164848.0 cr unu---uuuuu 161202e199606 xx s 000 0 eng 10.1093/hmg/5.6.809 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.6.809 FMRP is Associated to the Ribosomes Via RNA [Elektronische Daten] [Filippo Tamanini, Nicolle Meijer, Coleta Verheij, Patrick J. Willems, Hans Galjaard, Ben A. Oostra, André T. Hoogeveen] The FMR1 transcript is alternatively spliced and generates different splice variants coding for FMR1 proteins (FMRP) with a predicted molecular mass of 70-80 kDa. FMRP is widely expressed and localized in the cytoplasm. To study a possible interaction with other cellular components, FMRP was isolated and characterized under non-denaturing conditions. Under physiological salt conditions FMRP appears to have a molecular mass of >600 kDa, indicating a binding to other cellular components. This interaction is disrupted in the presence of high salt concentrations. The dissociation conditions to free FMRP from the complex are similar to the dissociation of FMRP from RNA as shown before. The binding of FMRP from the complex is also disrupted by RNAse treatment. That the association of FMRP to a high molecular weight complex possibly occurs via RNA, is further supported by the observation that the binding of FMRP, containing an Ile304Asn substitution, to the high molecular weight complex is reduced. An equal reduced binding of mutated FMRP to RNA in vitro was observed before under the same conditions. The reduced binding of FMRP with the Ile304Asn substitution further indicates that the interaction to the complex indeed occurs via FMRP and not via other RNA binding proteins. In a reconstitution experiment where the low molecular mass FMRP (70-80 kDa) is mixed with a reticulocyte lysate (enriched in ribosomes) it was shown that FMRP can associate to ribosomes and that this binding most likely occurs via RNA. © 1996 Oxford University Press Tamanini Filippo MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut Meijer Nicolle MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut Verheij Coleta MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut Willems Patrick J. Department of Medical Genetics, University of Antwerp, Antwerp, Belgium aut Galjaard Hans MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut Oostra Ben A. MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut Hoogeveen André T. MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut Human Molecular Genetics Oxford University Press 5/6(1996-06), 809-813 0964-6906 5:6<809 1996 5 hmg https://doi.org/10.1093/hmg/5.6.809 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.6.809 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tamanini Filippo MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut NATIONALLICENCE 700 1- Meijer Nicolle MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut NATIONALLICENCE 700 1- Verheij Coleta MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut NATIONALLICENCE 700 1- Willems Patrick J. Department of Medical Genetics, University of Antwerp, Antwerp, Belgium aut NATIONALLICENCE 700 1- Galjaard Hans MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut NATIONALLICENCE 700 1- Oostra Ben A. MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut NATIONALLICENCE 700 1- Hoogeveen André T. MGC-Department of Clinical Genetics, Erasmus University, Rotterdam, The Netherlands aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/6(1996-06), 809-813 0964-6906 5:6<809 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford