Acta Crystallographica Section D

Biological Crystallography

Volume 58, Part 9 (September 2002)

crystallization papers

Acta Cryst. (2002). D58, 1497-1500 [ doi:10.1107/S0907444902011770 ]

Purification, crystallization and initial crystallographic analysis of RNA polymerase holoenzyme from Thermus thermophilus

M. N. Vassylyeva, J. Lee, S. Sekine, O. Laptenko, S. Kuramitsu, T. Shibata, Y. Inoue, S. Borukhov, D. G. Vassylyev and S. Yokoyama

Abstract: RNA polymerase holoenzyme from Thermus thermophilus, consisting of six protein subunits ( [alpha] ₂, [beta] , [beta] ', [omega] and [sigma] ⁷⁰) and having a total molecular mass of about 450 kDa, was purified and crystallized by the hanging-drop vapour-diffusion technique under mild near-physiological conditions. The crystals diffract beyond 3 Å resolution. Careful analysis of diffraction data revealed that the crystals belong to space group P3₂, with unit-cell parameters a = b = 236.35, c = 249.04 Å, and have perfect twinning along the threefold axis. A complete data set at 3 Å resolution was collected and an unambiguous molecular-replacement solution was found using the structure of T. aquaticus RNA polymerase core enzyme as a search model. The refinement of structure and model building of the [sigma] ⁷⁰ subunit is now in progress.

Keywords: RNA polymerase.