Purification, crystallization and preliminary X-ray diffraction analysis of the histone chaperone cia1 from fission yeast.
In fission yeast, cia1+ is an essential gene that encodes a histone chaperone, a homologue of human CIA (CCG1-interacting factor A) and budding yeast Asf1p (anti-silencing function-1), which both facilitate nucleosome assembly by interacting with the core histones H3/H4. The conserved domain (residues 1-161) of the cia1+-encoded protein was expressed in Escherichia coli, purified to near-homogeneity and crystallized by the sitting-drop vapour-diffusion method. The protein was crystallized in the monoclinic space group C2, with unit-cell parameters a = 79.16, b = 40.53, c = 69.79 A, beta = 115.93 degrees and one molecule per asymmetric unit. The crystal diffracted to beyond 2.10 A resolution using synchrotron radiation.
Umehara
Takashi
T
RIKEN Genomic Sciences Center, 1-7-22 Suehiro, Tsurumi, Yokohama 230-0045, Japan.
Otta
Yumi
Y
Tsuganezawa
Keiko
K
Matsumoto
Takehisa
T
Tanaka
Akiko
A
Horikoshi
Masami
M
Padmanabhan
Balasundaram
B
Yokoyama
Shigeyuki
S
D018797:Cell Cycle Proteins / Q000737:chemistry*; D018360:Crystallography, X-Ray; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D020868:Gene Silencing; D005822:Genetic Vectors; D006657:Histones / Q000737:chemistry; D015233:Models, Statistical; D009707:Nucleosomes / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*; D012568:Schizosaccharomyces; D013696:Temperature; D014158:Transcription, Genetic; D014961:X-Ray Diffraction
D018797:Cell Cycle Proteins; C472823:Cia1p protein, S cerevisiae; D018076:DNA, Complementary; D006657:Histones; D009707:Nucleosomes; D029701:Saccharomyces cerevisiae Proteins