Open AccessCrystallization and preliminary X‐ray crystallographic analysis of sterol transcription factor Upc2 from Saccharomyces cerevisiae
Author(s) -
Ha Subin,
Tong Junsen,
Yang Huiseon,
Youn HyungSeop,
Eom Soo Hyun,
Im Young Jun
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112051597
Subject(s) - saccharomyces cerevisiae , crystallization , transcription factor , crystallography , chemistry , sterol , transcription (linguistics) , biochemistry , yeast , gene , cholesterol , organic chemistry , linguistics , philosophy
Upc2, a zinc‐cluster transcription factor, is a regulator of ergosterol biosynthesis in yeast. In response to sterol levels, the transcriptional activity of Upc2 is controlled by the C‐terminal domain. In this study, the C‐terminal regulatory domain of Upc2 from Saccharomyces cerevisiae was purified and crystallized by the vapour‐diffusion method. To improve the diffraction quality of Upc2 crystals, a Upc2 fusion protein in which 11 residues of the variable loop (residues 715–725) were replaced by T4 lysozymes in Upc2 (Upc2‐T4L) was engineered. The Upc2‐T4L crystals diffracted to 2.9 Å resolution using synchrotron radiation. The crystal was trigonal, belonging to space group P 3 2 with unit‐cell parameters a = 67.2, b = 67.2, c = 257.5 Å. The Matthews coefficient was determined to be 3.41 Å 3 Da −1 with two molecules in the asymmetric unit. Initial attempts to solve the structure by the single‐anomalous dispersion technique using selenomethionine were successful.