A conserved PUF-Ago-eEF1A complex attenuates translation elongation.
PUF (Pumilio/FBF) RNA-binding proteins and Argonaute (Ago) miRNA-binding proteins regulate mRNAs post-transcriptionally, each acting through similar, yet distinct, mechanisms. Here, we report that PUF and Ago proteins can also function together in a complex with a core translation elongation factor, eEF1A, to repress translation elongation. Both nematode (Caenorhabditis elegans) and mammalian PUF-Ago-eEF1A complexes were identified, using coimmunoprecipitation and recombinant protein assays. Nematode CSR-1 (Ago) promoted repression of FBF (PUF) target mRNAs in in vivo assays, and the FBF-1-CSR-1 heterodimer inhibited EFT-3 (eEF1A) GTPase activity in vitro. Mammalian PUM2-Ago-eEF1A inhibited translation of nonadenylated and polyadenylated reporter mRNAs in vitro. This repression occurred after translation initiation and led to ribosome accumulation within the open reading frame, roughly at the site where the nascent polypeptide emerged from the ribosomal exit tunnel. Together, these data suggest that a conserved PUF-Ago-eEF1A complex attenuates translation elongation.
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Kyle
K
Friend
Zachary T
ZT
Campbell
Amy
A
Cooke
Peggy
P
Kroll-Conner
Marvin P
MP
Wickens
Judith
J
Kimble
D000818:Animals; D060565:Argonaute Proteins / Q000378:metabolism*; D017173:Caenorhabditis elegans / Q000502:physiology; D029742:Caenorhabditis elegans Proteins / Q000378:metabolism; D002460:Cell Line; D039642:Eukaryotic Initiation Factors / Q000378:metabolism*; D006801:Humans; D020648:Peptide Elongation Factor 1 / Q000378:metabolism*; D014176:Protein Biosynthesis; D016601:RNA-Binding Proteins / Q000378:metabolism*
C115848:AGO1 protein, human; C466422:AGO2 protein, human; C517143:AGO3 protein, human; D060565:Argonaute Proteins; C544459:CSR-1 protein, C elegans; D029742:Caenorhabditis elegans Proteins; D039642:Eukaryotic Initiation Factors; C471243:PUM2 protein, human; D020648:Peptide Elongation Factor 1; D016601:RNA-Binding Proteins; C109286:fem-3-binding protein, C elegans