title: Solution structure of human P1•P2 heterodimer provides insights into the role of eukaryotic stalk in recruiting the ribosome-inactivating protein trichosanthin to the ribosome.
abstract: Lateral ribosomal stalk is responsible for binding and recruiting translation factors during protein synthesis. The eukaryotic stalk consists of one P0 protein with two copies of P1•P2 heterodimers to form a P0(P1•P2)₂ pentameric P-complex. Here, we have solved the structure of full-length P1•P2 by nuclear magnetic resonance spectroscopy. P1 and P2 dimerize via their helical N-terminal domains, whereas the C-terminal tails of P1•P2 are unstructured and can extend up to ∼125 Å away from the dimerization domains. (15)N relaxation study reveals that the C-terminal tails are flexible, having a much faster internal mobility than the N-terminal domains. Replacement of prokaryotic L10(L7/L12)₄/L11 by eukaryotic P0(P1•P2)₂/eL12 rendered Escherichia coli ribosome, which is insensitive to trichosanthin (TCS), susceptible to depurination by TCS and the C-terminal tail was found to be responsible for this depurination. Truncation and insertion studies showed that depurination of hybrid ribosome is dependent on the length of the proline-alanine rich hinge region within the C-terminal tail. All together, we propose a model that recruitment of TCS to the sarcin-ricin loop required the flexible C-terminal tail, and the proline-alanine rich hinge region lengthens this C-terminal tail, allowing the tail to sweep around the ribosome to recruit TCS.
authors.affiliation: School of Life Sciences, Centre for Protein Science and Crystallography, The Chinese University of Hong Kong, Shatin, Hong Kong, China and Department of Biology, Faculty of Science, Niigata University, Ikarashi 2-8050, Nishi-ku, Niigata 950-2181, Japan.
authors.forename: Ka-Ming
authors.initials: KM
authors.lastname: Lee
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Yusa
authors.forename: Lai-On
authors.initials: LO
authors.lastname: Chu
authors.forename: Conny Wing-Heng
authors.initials: CW
authors.lastname: Yu
authors.forename: Moe
authors.initials: M
authors.lastname: Oono
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Miyoshi
authors.forename: Kosuke
authors.initials: K
authors.lastname: Ito
authors.forename: Pang-Chui
authors.initials: PC
authors.lastname: Shaw
authors.forename: Kam-Bo
authors.initials: KB
authors.lastname: Wong
authors.forename: Toshio
authors.initials: T
authors.lastname: Uchiumi
mesh_terms: D000225:Adenine / Q000378:metabolism; D004926:Escherichia coli; D006801:Humans; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010750:Phosphoproteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D012269:Ribosomal Proteins / Q000737:chemistry*; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism*; D015978:Trichosanthin / Q000378:metabolism*
chemical_list: D010750:Phosphoproteins; D012269:Ribosomal Proteins; C031937:phosphoprotein P2, ribosomal; C035690:ribosomal phosphoprotein P1; D015978:Trichosanthin; D000225:Adenine
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