title: Expression cloning and characterization of a novel gene that encodes the RNA-binding protein FAU-1 from Pyrococcus furiosus.
abstract: We systematically screened a genomic DNA library to identify proteins of the hyperthermophilic archaeon Pyrococcus furiosus using an expression cloning method. One gene product, which we named FAU-1 (P. furiosus AU-binding), demonstrated the strongest binding activity of all the genomic library-derived proteins tested against an AU-rich RNA sequence. The protein was purified to near homogeneity as a 54 kDa single polypeptide, and the gene locus corresponding to this FAU-1 activity was also sequenced. The FAU-1 gene encoded a 472-amino-acid protein that was characterized by highly charged domains consisting of both acidic and basic amino acids. The N-terminal half of the gene had a degree of similarity (25%) with RNase E from Escherichia coli. Five rounds of RNA-binding-site selection and footprinting analysis showed that the FAU-1 protein binds specifically to the AU-rich sequence in a loop region of a possible RNA ligand. Moreover, we demonstrated that the FAU-1 protein acts as an oligomer, and mainly as a trimer. These results showed that the FAU-1 protein is a novel heat-stable protein with an RNA loop-binding characteristic.
authors.affiliation: Doi Bioasymmetry Project, Exploratory Research for Advanced Technology (ERATO), Japan Science and Technology Corporation, 4-1-8 Kawaguchi, Saitama 332-0012, Japan. akio@sfc.keio.ac.jp
authors.forename: Akio
authors.initials: A
authors.lastname: Kanai
authors.forename: Hanako
authors.initials: H
authors.lastname: Oida
authors.forename: Nana
authors.initials: N
authors.lastname: Matsuura
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Doi
mesh_terms: D008969:Molecular Sequence Data; D011485:Protein Binding / Q000502:physiology; D020027:Pyrococcus furiosus / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D016601:RNA-Binding Proteins; D012313:RNA