Expression, crystallization and preliminary X-ray analysis of the Pyrococcus abyssi protein homologue of Saccharomyces cerevisiae Nip7p.

ABSTRACT

Saccharomyces cerevisiae Nip7p is a nucleolar protein required for accurate processing of the 27S precursor of the 25S and 5.8S ribosomal RNAs. Nip7p homologues are found in eukaryotes and archaea. The Pyrococcus abyssi homologue of Nip7p (PaNip7) was cloned, expressed in Escherichia coli and purified for crystallization. X-ray diffraction data were collected from native crystals and an iodide derivative using synchrotron radiation. PaNip7 native crystals diffract to 1.8 A and belong to space group C2, with unit-cell parameters a = 88.49, b = 90.28, c = 63.35 A, beta = 134.29 degrees. The PaNip7 structure was solved using the SIRAS method.