Purification, crystallization and preliminary X-ray diffraction analysis of yeast nucleosome-assembly factor Cia1p.
Yeast Cia1p is a homologue of human CIA (CCG1-interacting factor A), which possesses nucleosome-assembly activity and interacts with the human TFIID subunit CCG1 and the C-terminal domain of histone H3. The yeast Cia1p without the C-terminal polyanionic stretch has been expressed in Escherichia coli, purified to homogeneity and crystallized by the hanging-drop vapour-diffusion method using PEG 8000 as precipitant. The protein was crystallized in orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 106.70, b = 46.92, c = 40.60 A and one molecule in the asymmetric unit. The crystal diffracted beyond 2.95 A resolution using synchrotron radiation.
D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D009707:Nucleosomes / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000302:isolation & purification
D018797:Cell Cycle Proteins; C472823:Cia1p protein, S cerevisiae; D009707:Nucleosomes; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins