Crystallization and preliminary crystallographic studies of human RIG-I in complex with double-stranded RNA.
Retinoic acid inducible gene-I (RIG-I) is an essential component of the innate immune system that is responsible for the detection and elimination of invading viruses. RIG-I recognizes viral RNAs inside the cell and then initiates downstream signalling to activate the IRF-3 and NF-kappaB genes, which results in the production of type I interferons. RIG-I is composed of an N-terminal CARD domain for signalling and C-terminal helicase and repressor domains for RNA recognition. A RIG-I-RNA binding assay was performed to investigate the in vitro RIG-I-RNA binding properties. Selenomethionine-incorporated RIG-I was expressed using Escherichia coli and purified for crystallization. X-ray data were collected from RIG-I-dsRNA complex crystals to 2.8 A resolution using synchrotron radiation.
Moon
Hyunjin
H
Department of Biological Science, University of Seoul, 90 Cheonnong-dong, Dongdaemun-gu, Seoul 130-743, Republic of Korea.
Choe
Jungwoo
J
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003625:Data Collection; D004926:Escherichia coli / Q000235:genetics; D004996:Ethidium / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012330:RNA, Double-Stranded / Q000378:metabolism*; D011971:Receptors, Immunologic; D013223:Statistics as Topic; D017356:Synchrotrons; D013696:Temperature
D005456:Fluorescent Dyes; D012330:RNA, Double-Stranded; D011971:Receptors, Immunologic; D006639:Histidine; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases; D004996:Ethidium