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89 Multisubunit RNA polymerases Patrick Cramer Transcription of the genetic information in all cells is carried quantities of cell culture without the benefit of over- out by multisubunit RNA polymerases (RNAPs). Comparison expression. In both cases, interpretation of the experimental of the crystal structures of a bacterial and a eukaryotic RNAP electron density maps relied on the placement of known reveals a conserved core that comprises the active site and a subunit structures and phase combination. In the case of multifunctional clamp. Together with a further structure of bRNAP, X-ray diffraction was weak, radiation sensitive eukaryotic RNAP bound to DNA and RNA, these results and anisotropic. The interpretation of maps with weak elucidate many aspects of the transcription mechanism, sidechain electron density relied on noncrystallographic including initiation, elongation, nucleotide addition, processivity symmetry averaging and data from selenomethionyl and proofreading. bRNAP. In the case of yRNAP, a mutant yeast strain had to be used to produce Pol II lacking two substoichiometric Addresses subunits. Nonisomorphous and weakly diffracting initial Institute of Biochemistry, Gene Center, University of Munich, Pol II crystals were improved dramatically by a soaking Feodor-Lynen-Strasse 25, 81377 Munich, Germany; procedure that induced crystal shrinkage. Single heavy atoms e-mail: cramer@lmb.uni-muenchen.de could not be detected in Patterson maps and initial phasing Current Opinion in Structural Biology 2002, 12:89–97 therefore relied on heavy atom clusters. Single heavy atom deriv