Molecular mechanism of target RNA transcript recognition by Argonaute-guide complexes.
Argonaute proteins participate in conferring all known functions of RNA-mediated gene silencing phenomena. However, prior to structural investigations of this evolutionarily conserved family of proteins, there was little information concerning their mechanisms of action. Here, we describe our crystallographic analysis of the PIWI domain of an archaeal Argonaute homolog, AfPiwi. Our structural analysis revealed that the Argonaute PIWI fold incorporates both an RNase-H-like catalytic domain and an anchor site for the obligatory 5' phosphate of the RNA guide strand. RNA-AfPiwi binding assays combined with crystallographic studies demonstrated that AfPiwi interacts with RNA via a conserved region centered on the carboxyl terminus of the protein, utilizing a novel metal-binding site. A model of the PIWI domain of Argonaute in complex with a small interfering RNA (siRNA)-like duplex is consistent with much of the existing biochemical and genetic data, explaining the specificity of the RNA-directed RNA endonuclease reaction and the importance of the 5' region of microRNAs (miRNAs) (the "seed") to nucleate target RNA recognition and provide high-affinity guide-target interactions.
Parker
J S
JS
Section of Structural Biology, Institute of Cancer Research, Chester Beatty Laboratories, London, United Kingdom.
Roe
S M
SM
Barford
D
D
D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D019611:Archaeoglobus fulgidus / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D046911:Macromolecular Substances; D008954:Models, Biological; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D019642:RNA, Archaeal / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D034743:RNA-Induced Silencing Complex / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D058727:RNA, Small Untranslated
D019843:Archaeal Proteins; D046911:Macromolecular Substances; D019642:RNA, Archaeal; D034741:RNA, Small Interfering; D034743:RNA-Induced Silencing Complex; D058727:RNA, Small Untranslated