RNA polymerase II (Pol II)

RNA polymerase II (Pol II) is a 12 subunit protein complex from yeast to human that is required for gene expression. Gdown1 containing Pol II [Pol II (G)] is a special form of Pol II that is catalytically inactive and heavily depends on the 30-subunit Mediator complex for its activator and basal dependent function in vitro. Here we report for the first time, the identification and the generation of a 15-subunit human Mediator complex via the novel multibac baculovirus expression system that is fully responsive to Pol II (G).

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Minichromosomemaintenance (MCM) proteins are part of the replication licensing factor (RLF-M), which limits the initiationofDNAreplication to once per cell cycle.We have previously reported that higher order complexes of mam-malian pol II and general pol II transcription factors, referred to as pol II holoenzyme, also contain MCM pro-teins. In the present study we have analyzed in detail the interaction between MCM2 and pol II holoenzyme. N- and C- terminal deletions were introduced into epitope-tagged MCM2 and the truncated proteins were transiently expressed in 293 cells. ...

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RNA polymerase II (pol II) produces all mRNA in eukaryotic cells. Structures of the ten-subunit core of yeast pol II in free form [1, 2], in complex with DNA and RNA [3], and in complex with alpha-amanitin [4] have elucidated the mRNA transcription mech-anism [reviewed in 5, 6]. Based on this work, models for the com-plete 12-subunit pol II were obtained [7, 8]. We have subsequently solved the first structure of pol II in complex with a transcription factor, the elongation factor TFIIS [9, 10].

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In the first half of the talk, the current state of our ongoing structure-function analysis of the mRNA transcription cycle will be dis-cussed. The crystallographic structures of the complete 12-subunit RNA polymerase II in free form and with bound DNA and RNA have been determined as atomic models. The structure of the complete Pol II elongation complex bound by the transcript cleavage factor TFIIS explained how Pol II uses a single tunable active site for both RNA synthesis and RNA cleavage.

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Symposia Lectures Monday Monday, July 9, 2007 A2-L1 J. Diffley No abstract available. A2-L2 Molecular bioimaging of gene transcription P. Cramer Gene Center, Munich, GERMANY In the first half of the talk, the current state of our ongoing structure-function analysis of the mRNA transcription cycle will be discussed. The crystallographic structures of the complete 12-subunit RNA polymerase II in free form and with bound DNA and RNA have been determined as atomic models.

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