Crystal structures of restrictocin-inhibitor complexes with implications for RNA recognition and base flipping.
The cytotoxin sarcin disrupts elongation factor binding and protein synthesis by specifically cleaving one phosphodiester bond in ribosomes. To elucidate the molecular basis of toxin action, we determined three cocrystal structures of the sarcin homolog restrictocin bound to different analogs that mimic the target sarcin/ricin loop (SRL) structure of the rat 28S rRNA. In these structures, restrictocin contacts the bulged-G motif and an unfolded form of the tetraloop of the SRL RNA. In one structure, toxin loops guide selection of the target site by contacting the base critical for recognition (G4319) and the surrounding S-shaped backbone. In another structure, base flipping of the tetraloop enables cleavage by placing the target nucleotide in the active site with the nucleophile nearly inline for attack on the scissile bond. These structures provide the first views of how a site-specific protein endonuclease recognizes and cleaves a folded RNA substrate.
Yang
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Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, Illinois 60637 USA.
Gérczei
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Glover
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Correll
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D000485:Allergens; D000818:Animals; D052179:Antigens, Plant; D001483:Base Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D005656:Fungal Proteins / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000378:metabolism*; D012339:RNA, Ribosomal, 28S / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D012260:Ribonucleases / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity
C081947:ASPF1 protein, Aspergillus fumigatus; D000485:Allergens; D052179:Antigens, Plant; C105968:C-SAR protein, Aspergillus clavatus; D005656:Fungal Proteins; D009841:Oligonucleotides; D011500:Protein Synthesis Inhibitors; D012339:RNA, Ribosomal, 28S; D016601:RNA-Binding Proteins; C036196:MITF protein, Aspergillus restrictus; D012260:Ribonucleases