rDNA Transcription

Ribosomes, the cellular factories for protein production, are among the most intricate and complex ribonucleoprotein (RNP)-assemblies in the cell. Their biogenesis is tightly regulated and requires the coordinated activity of many cellular processes and can therefore be considered as a well-suited model system to study mechanisms of biogenesis of RNP complexes. Synthesis of the majority of ribosomal RNAs in eukaryotes is executed by a highly specialized RNA polymerase, Pol I. Our major interests are to understand mechanisms and regulation of the RNA polymerase I transcription machinery and to study how rRNA synthesis is coupled to rRNA maturation and assembly of pre-ribosomes.

Our goals are to reconstitute transcription initiation, elongation and termination from a minimal transcription system, to understand the roles of single subunits of the transcription machinery and to compare features of Pol I transcription to other nuclear RNA polymerases.

Selected publications

Griesenbeck, J., Tschochner, H., and Grohmann, D. (2017). Structure and Function of RNA Polymerases and the Transcription Machineries. Subcell. Biochem. 83, 225–270.

Pilsl, M., Crucifix, C., Papai, G., Krupp, F., Steinbauer, R., Griesenbeck, J., Milkereit, P., Tschochner, H., and Schultz, P. (2016b). Structure of the initiation-competent RNA polymerase I and its implication for transcription. Nat. Commun. 7, 12126.

Pilsl, M., Merkl, P.E., Milkereit, P., Griesenbeck, J., and Tschochner, H. (2016a). Analysis of S. cerevisiae RNA Polymerase I Transcription In Vitro. Methods Mol. Biol. Clifton NJ 1455, 99–108.

Merkl, P., Perez-Fernandez, J., Pilsl, M., Reiter, A., Williams, L., Gerber, J., Böhm, M., Deutzmann, R., Griesenbeck, J., Milkereit, P., and Tschochner H. (2014). Binding of the termination factor Nsi1 to its cognate DNA site is sufficient to terminate RNA polymerase I transcription in vitro and to induce termination in vivo. Mol. Cell. Biol. 34, 3817–3827.

Németh, A., Perez-Fernandez, J., Merkl, P., Hamperl, S., Gerber, J., Griesenbeck, J., and Tschochner, H. (2013). RNA polymerase I termination: Where is the end? Biochim. Biophys. Acta 1829, 306–317.

Reiter, A., Hamperl, S., Seitz, H., Merkl, P., Perez-Fernandez, J., Williams, L., Gerber, J., Németh, A., Léger, I., Gadal, O., Milkereit, P., Griesenbeck, J., and Tschochner, H. (2012). The Reb1-homologue Ydr026c/Nsi1 is required for efficient RNA polymerase I termination in yeast. EMBO J. 31, 3480–3493.

Reiter A, Steinbauer R, Philippi A, Gerber J, Tschochner H, Milkereit P, and Griesenbeck J. (2011). Reduction in Ribosomal Protein Synthesis Is Sufficient To Explain Major Effects on Ribosome Production after Short-Term TOR Inactivation in Saccharomyces cerevisiae. Mol Cell Biol. 2011 Feb;31(4):803-17.

Philippi, A., Steinbauer R., Reiter A., Fath S., Leger-Silvestre I., Milkereit P., Griesenbeck J. and Tschochner H. (2010) TOR-dependent reduction in the expression level of Rrn3p lowers the activity of the yeast RNA Pol I machinery, but does not account for the strong inhibition of rRNA production. Nucleic Acids Research, 2010, 38 (16), 5315-5326.

Reich, C., Zeller, M., Milkereit, P., Hausner, W., Cramer, P., Tschochner, H. and Thomm, M. (2009)  The archaeal RNA polymerase subunit P and the eukaryotic polymerase subunit Rpb12 are interchangeable in vivo and in vitro. Mol Microbiology 71, 989-1002. 

Clemente-Blanco, A., Mayán-Santos, M., Schneider, D.A., Machín, F., Jarmuz, A., Tschochner, H. and Aragón, L. (2009) Cdc14 inhibits transcription by RNA polymerase I during anaphase. Nature, 458, 219-222. 

Gerber, J., Reiter, A., Steinbauer, R., Jakob, S., Kuhn, C.-D., Cramer, P., Griesenbeck, J., Milkereit, P. and Tschochner, H. (2008) Site specific phosphorylation of yeast RNA polymerase I. Nucleic Acid Research, 36, 793-802.

Kuhn, C.-D., Geiger, S., Baumli, S., Gartmann, M., Gerber, J., Jennebach, S., Mielke, T., Tschochner, H., Beckmann, R. and Cramer, P. (2007) Functional architecture of yeast RNA polymerase I. Cell, 131, 1260-1272.

Fath, S., Kobor, M., Philippi, A., Greenblatt, M. and Tschochner, H. (2004) Dephosphorylation of RNA polymerase I by Fcp1p is required for efficient rRNA synthesis J. Biol. Chem. 279(24), 25251-9. 

Bier, M., Fath S. and Tschochner, H. (2004) The composition of the RNA polymerase I transcription machinery switches from initiation to elongation mode FEBS Lett. 564 (1-2), 41-46.

Bischler, N., Brino, L., Carles, C., Riva, M., Tschochner, H., Mallouh, V. and Schultz, P. (2002) Localisation of the yeast RNA polymerase I-specific subunits. EMBO J. 21, 4136-4144.

Iben, S., Tschochner, H., Bier, M.,  Hoogstraten, D., Hozak, P., Egly, J.-M. and Grummt, I. (2002) TFIIH plays an essential role in RNA polymerase I  transcription Cell 109,  297-306 

Fath, S., Milkereit, P., Peyroche, G., Riva, M., Carles, Ch. and Tschochner, H.  (2001) Differential roles of phosphorylation in the formation of transcriptional active RNA polymerase I. Proc. Natl. Acad. Sci. USA 98, 14334-14339. 

Peyroche, G., Milkereit, P., Carles, C., Tschochner, H.,  Schultz, P., Sentenac, A. and Riva, M. (2000) The recruitment of RNA polymerase I on rDNA is mediated by the interaction of the A43 subunit with Rrn3. EMBO J. 19(20):5473-5482.

Fath, S., Milkereit, P., Podtelejnikov, A., Bischler, N., Schultz, P., Mann, M. and Tschochner, H. (2000) Association of yeast RNA polymerase I with a nucleolar substructure active in rRNA synthesis and processing J. Cell Biol. 149, 575-589.   

Milkereit, P. and Tschochner H. (1998) A specialized form of RNA polymerase I, essential for initiation and growth dependent regulation of rRNA synthesis, is disrupted during transcription EMBO J. 17, 3692-3703. 

Bischler, N., Balavoine, F., Milkereit, P., Tschochner, H., Mioskowski, C. and Schultz, P. (1998) Specific interaction and two-dimensional crystallisation of histidine tagged yeast RNA polymerase I on Nickel-chelating lipids  Biophys. J. 74, 1522-1532.

Milkereit, P., Schultz, P. and Tschochner, H. (1997) Resolution of RNA polymerase I into dimers and monomers and their function in transcription  Biol. Chem. 378, 1433-144.

Tschochner, H. and Milkereit, P. (1997) RNA polymerase I from S. cerevisiae depends on an additional factor to release terminated transcripts from the template. FEBS Lett. 410, 461-466.

Tschochner, H. (1996) A novel RNA polymerase I-dependent RNase activity that shortens nascent transcripts from the 3´end. Proc. Natl. Acad. Sci. USA 93, 12914-12919.

Wettach, J., Gohl, H.P., Tschochner, H. and Thomm, M. (1995) Functional interaction of yeast and human TATA-binding protein with an archael RNA polymerase and promoter. Proc. Natl. Acad. Sci. U.S.A. 92, 472-476.

All relevant publications in PubMed.

• since July 2011

  DFG collaborative research Center SFB960 at the University of Regensburg: Ribosome formation: principles of RNP biogenesis and control of their function

  Project A1: Analyses of the RNA polymerase I machinery: specific transcriptional mechanisms and comparison with other RNA polymerases

Christophe Carles, CEA Saclay, Gif-sur-Yvette, France.

Olivier Gadal, Laboratoire de Biologie Moléculaire Eucaryote, Université Paul Sabatier, Toulouse III, France.

Thomas Moss, Centre de Recherche en Cancérologie, Université Laval, Quebec, Canada.

Patrick Schultz, Institut de Génétique et de Biologie Moléculaire et Cellulaire, Illkirch, France.