title: Construction of the 'minimal' SRP that interacts with the translating ribosome but not with specific membrane receptors in Escherichia coli.
abstract: Escherichia coli signal recognition particle (SRP) consists of 4.5S RNA and Ffh protein. In contrast to eukaryotes, it remains unclear whether translation arrest takes place in prokaryotic cells. To study this problem we constructed a fusion of the M domain of Ffh protein with a cleavable affinity tag. This mutant Ffh, in a complex with 4.5S RNA, can bind signal peptide at the translating ribosome but is unable to bind the membrane. This SRP-ribosome complex should accumulate in the cell if translation is arrested. To test this, the complex was purified from the cells by ultracentrifugation and affinity chromatography. The composition of the complex was analyzed and found to consist of ribosomal RNAs and proteins, the Ffh M domain and 4.5S RNA. The accumulation of this complex in the cell in significant amounts indicated that SRP-mediated translation arrest did occur in bacterial cells.
authors.affiliation: Department of Chemistry, Moscow State University, 119899, Moscow, Russia.
authors.forename: Olga N
authors.initials: ON
authors.lastname: Avdeeva
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Myasnikov
authors.forename: Petr V
authors.initials: PV
authors.lastname: Sergiev
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Bogdanov
authors.forename: Richard
authors.initials: R
authors.lastname: Brimacombe
authors.forename: Olga A
authors.initials: OA
authors.lastname: Dontsova
mesh_terms: D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism; D012329:RNA, Bacterial; D012335:RNA, Ribosomal / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D018271:Signal Recognition Particle / Q000378:metabolism*
chemical_list: C409968:4.5S RNA; D029968:Escherichia coli Proteins; C082931:Ffh protein, E coli; D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D011994:Recombinant Proteins; D018271:Signal Recognition Particle