title: An archaeal protein homologous to mammalian SRP54 and bacterial Ffh recognizes a highly conserved region of SRP RNA.
abstract: The gene encoding the 54 kDa protein of signal recognition particle (SRP54) in the hyperthermophilic archaeon Pyrococcus furiosus has been cloned and sequenced. Recombinant P. furiosus SRP54 (pf-SRP54) and the N-terminal G-domain and C-terminal M-domain (pf-SRP54M) of pf-SRP54 with an amino-terminal addition of six histidine residues were expressed in Escherichia coli and subjected to binding experiments for SRP RNA, non-conserved 213-nucleotide RNA (helices 1, 2, 3, 4 and 5) and conserved 107-nucleotide RNA (helices 6 and 8) from SRP RNA. The RNA binding properties of the purified protein were determined by filter binding assays. The histidine-tagged pf-SRP54M bound specifically to the conserved 107-nucleotide RNA in the absence of pf-SRP19, unlike the eukaryotic homologue, with an apparent binding constant (K) of 18 nM.
authors.affiliation: School of Bioresources, Hiroshima Prefectural University, Shobara City, 727-0023, Hiroshima, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Maeshima
authors.forename: E
authors.initials: E
authors.lastname: Okuno
authors.forename: T
authors.initials: T
authors.lastname: Aimi
authors.forename: T
authors.initials: T
authors.lastname: Morinaga
authors.forename: T
authors.initials: T
authors.lastname: Itoh
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019843:Archaeal Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D017124:Conserved Sequence; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008322:Mammals; D008715:Methionine; D008969:Molecular Sequence Data; D020027:Pyrococcus furiosus / Q000235:genetics*; D012313:RNA / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D018271:Signal Recognition Particle / Q000235:genetics* / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D029968:Escherichia coli Proteins; C082931:Ffh protein, E coli; C117074:SRP19 protein, human; C069268:SRP54 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D018271:Signal Recognition Particle; D012313:RNA; D008715:Methionine