caa a22 4500 469090766 CHVBK 20180323133016.0 cr unu---uuuuu 170328e19920901xx s 000 0 eng 10.1007/BF00419662 doi (NATIONALLICENCE)springer-10.1007/BF00419662 In vitro reconstitution of U1 and U2 snRNPs from isolated proteins and snRNA [Elektronische Daten] [V. Sumpter, A. Kahrs, U. Fischer, U. Kornstädt, R. Lührmann] In this paper we describe a method for preparing native, RNA-free, proteins from anti-m3G purified snRNPs (U1, U2, U4/U6 and U5) and the subsequent quantitative reconstitution of U1 and U2 snRNPs from purified proteins and snRNA. Reconstituted U1 and U2 snRNPs contained the full complement of core proteins, B, B′, D1, D2, D3, E, F and G. Both the U1 and U2 reconstituted particles were stable in CsCl gradients and had the expected buoyant density of 1.4 g/cm3. Reconstituted RNP particle formation was not competited by a 50 fold molar excess of tRNA, as determined by gel retardation assays. However, U1 and U2 particle formation was reduced in the presence of an excess of cold U1 or U2 snRNA demonstrating a specific RNA-protein interaction. U1 and U2 snRNPs were also efficiently reconstituted in vitro, utilizing proteins prepared from mono Q purified U1 and U2 snRNPs. This suggests that for the assembly of snRNPs in vitro no auxiliary proteins other than bona fide snRNP proteins appear to be required. The potential of this reconstitution technique for investigating snRNP assembly and snRNA-protein interactions is discussed. Kluwer Academic Publishers, 1992 U snRNPs nationallicence in vitro reconstitution nationallicence RNA-protein interaction nationallicence PEG : Polyethelene glycol nationallicence PMSF : Phenylmethyl sulfonylfluoride nationallicence TP : total proteins nationallicence mAb : monoclonal antibody nationallicence Sumpter V. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut Kahrs A. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut Fischer U. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut Kornstädt U. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut Lührmann R. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut Molecular Biology Reports Kluwer Academic Publishers 16/4(1992-09-01), 229-240 0301-4851 16:4<229 1992 16 11033 https://doi.org/10.1007/BF00419662 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00419662 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sumpter V. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut NATIONALLICENCE 700 1- Kahrs A. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut NATIONALLICENCE 700 1- Fischer U. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut NATIONALLICENCE 700 1- Kornstädt U. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut NATIONALLICENCE 700 1- Lührmann R. Institut für Molekularbiologie und Tumorforschung, der Philipps-Universität Marburg, Emil-Mannkopff-Straße 2, D-3550, Marburg, Germany aut NATIONALLICENCE 773 0- Molecular Biology Reports Kluwer Academic Publishers 16/4(1992-09-01), 229-240 0301-4851 16:4<229 1992 16 11033 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer