MSMEG_6292, a Mycobacterium smegmatis RNA polymerase secondary channel-binding protein: purification, crystallization and X-ray diffraction analysis

Joseph, Abyson and Nagaraja, Valakunja and Natesh, Ramanathan (2018) MSMEG_6292, a Mycobacterium smegmatis RNA polymerase secondary channel-binding protein: purification, crystallization and X-ray diffraction analysis. In: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, 74 (9). pp. 543-548.

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Abstract

The transcriptional activity of RNA polymerase (RNAP) is controlled by a diverse set of regulatory factors. A subset of these regulators modulate the activity of RNAP through its secondary channel. Gre factors reactivate stalled elongation complexes by enhancing the intrinsic cleavage activity of RNAP. In the present study, the protein MSMEG_6292, a Gre-factor homologue from Mycobacterium smegmatis, was expressed heterologously in Escherichia coli and purified using standard chromatographic techniques. The hanging-drop vapour-diffusion crystallization method yielded diffraction-quality crystals. The crystals belonged to the trigonal space group P3(1)21 (or its enantiomorph P3(2)21), with unit-cell parameters a = b = 83.15, c = 107.07 angstrom, alpha = beta = 90, gamma = 120 degrees. The crystals diffracted to better than 3.0 angstrom resolution. Molecular-replacement attempts did not yield any phasing models; hence, platinum derivatization was carried out with K2PtCl4 and derivative data were collected to 3.4 angstrom resolution.

Item Type:	Journal Article
Publication:	ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS
Publisher:	INT UNION CRYSTALLOGRAPHY
Additional Information:	Copy right for this article belong to INT UNION CRYSTALLOGRAPHY, 2 ABBEY SQ, CHESTER, CH1 2HU, ENGLAND
Keywords:	transcription regulation; RNA polymerase; Gre-factor homologue; mycobacteria; X-ray diffraction; Mycobacterium smegmatis; MSMEG_6292
Department/Centre:	Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited:	26 Sep 2018 15:27
Last Modified:	26 Sep 2018 15:27
URI:	http://eprints.iisc.ac.in/id/eprint/60739

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