Ohmayer, Uli and Perez-Fernandez, Jorge and Hierlmeier, Thomas and Pöll, Gisela and Williams, Lydia and Griesenbeck, Joachim and Tschochner, Herbert and Milkereit, Philipp (2012) Local Tertiary Structure Probing of Ribonucleoprotein Particles by Nuclease Fusion Proteins. Plos One 7, e42449.
|License: Creative Commons Attribution|
PDF - Published Version
Analyses of the conformational dynamics of the numerous cellular ribonucleoprotein particles (RNP) significantly contribute to the understanding of their modes of action. Here, we tested whether ribonuclease fusion proteins incorporated into RNPs can be used as molecular probes to characterize the local RNA environment of these proteins. Fusion proteins of micrococcal nuclease (MNase) with ribosomal proteins were expressed in S. cerevisae to produce in vivo recombinant ribosomes which have a ribonuclease tethered to specific sites. Activation of the MNase activity by addition of calcium led to specific rRNA cleavage events in proximity to the ribosomal binding sites of the fusion proteins. The dimensions of the RNP environment which could be probed by this approach varied with the size of the linker sequence between MNase and the fused protein. Advantages and disadvantages of the use of MNase fusion proteins for local tertiary structure probing of RNPs as well as alternative applications for this type of approach in RNP research are discussed.
|Date:||2 August 2012|
|Institutions:||Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Prof. Dr. Herbert Tschochner|
Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Dr. Joachim Griesenbeck
Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III > Dr. Philipp Milkereit
|Projects:||Open Access Publizieren (DFG)|
|Subjects:||500 Science > 570 Life sciences|
600 Technology > 610 Medical sciences Medicine
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||Yes|
|Deposited On:||07 Sep 2012 06:58|
|Last Modified:||13 Mar 2014 19:12|