Cloning, purification, crystallization and preliminary X-ray crystallographic analysis of the N-terminal domain of DEAD-box RNA helicase fromStaphylococcus aureusstrain Mu50

Abstract

DEAD-box helicases are enzymes with an ATP-dependent RNA-unwinding function that are involved in a variety of cellular processes including RNA splicing, ribosome biogenesis and RNA degradation. In this study, the N-terminal domain of DEAD-box RNA helicase from Staphylococcus aureus strain Mu50 was overexpressed in Escherichia coli, purified and crystallized. Diffraction data were collected to 2.60 Å resolution using a synchrotronradiation source. The crystal belonged to space group P1, with unit-cell parameters a = 70. 81, b = 80.23, c = 86.25 Å , = 69.54, = 66.54, = 87.32 . The unit cell contained six molecules, with a corresponding V M of 2.91 Å 3 Da À1 and a solvent content of 56.1%.

Dependence of RIG-I Nucleic Acid-Binding and ATP Hydrolysis on Activation of Type I Interferon Response

“…The cells were then further incubated at 16℃ for 24 h. After harvesting, resuspended cells were disrupted by sonication. All recombinant proteins were purified with Ni fast protein liquid chromatography (FPLC) using standard protocols (GE Healthcare, Piscataway, NJ, USA) ( 17 ).…”