title: Turning catalytically inactive human Argonaute proteins into active slicer enzymes.
abstract: Argonaute proteins interact with small RNAs that guide them to complementary target RNAs, thus leading to inhibition of gene expression. Some but not all Argonaute proteins are endonucleases and can cleave the complementary target RNA. Here, we have mutated inactive human Ago1 and Ago3 and generated catalytic Argonaute proteins. We find that two short sequence elements at the N terminus are important for activity. In addition, PIWI-domain mutations in Ago1 may misarrange the catalytic center. Our work helps in understanding of the structural requirements that make an Argonaute protein an active endonucleolytic enzyme.
authors.affiliation: Biochemistry Center Regensburg, Laboratory for RNA Biology, University of Regensburg, Germany.
authors.forename: Judith
authors.initials: J
authors.lastname: Hauptmann
authors.forename: Anne
authors.initials: A
authors.lastname: Dueck
authors.forename: Simone
authors.initials: S
authors.lastname: Harlander
authors.forename: Janina
authors.initials: J
authors.lastname: Pfaff
authors.forename: Rainer
authors.initials: R
authors.lastname: Merkl
authors.forename: Gunter
authors.initials: G
authors.lastname: Meister
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D060565:Argonaute Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002384:Catalysis; D039642:Eukaryotic Initiation Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D012323:RNA Processing, Post-Transcriptional; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
chemical_list: C115848:AGO1 protein, human; C466422:AGO2 protein, human; C517143:AGO3 protein, human; C517144:AGO4 protein, human; D060565:Argonaute Proteins; D039642:Eukaryotic Initiation Factors; D035683:MicroRNAs; D011993:Recombinant Fusion Proteins