title: Translation initiation on mRNAs bound by nuclear cap-binding protein complex CBP80/20 requires interaction between CBP80/20-dependent translation initiation factor and eukaryotic translation initiation factor 3g.
abstract: In the cytoplasm of mammalian cells, either cap-binding proteins 80 and 20 (CBP80/20) or eukaryotic translation initiation factor (eIF) 4E can direct the initiation of translation. Although the recruitment of ribosomes to mRNAs during eIF4E-dependent translation (ET) is well characterized, the molecular mechanism for CBP80/20-dependent translation (CT) remains obscure. Here, we show that CBP80/20-dependent translation initiation factor (CTIF), which has been shown to be preferentially involved in CT but not ET, specifically interacts with eIF3g, a component of the eIF3 complex involved in ribosome recruitment. By interacting with eIF3g, CTIF serves as an adaptor protein to bridge the CBP80/20 and the eIF3 complex, leading to efficient ribosome recruitment during CT. Accordingly, down-regulation of CTIF using a small interfering RNA causes a redistribution of CBP80 from polysome fractions to subpolysome fractions, without significant consequence to eIF4E distribution. In addition, down-regulation of eIF3g inhibits the efficiency of nonsense-mediated mRNA decay, which is tightly coupled to CT but not to ET. Moreover, the artificial tethering of CTIF to an intercistronic region of dicistronic mRNA results in translation of the downstream cistron in an eIF3-dependent manner. These findings support the idea that CT mechanistically differs from ET.
authors.affiliation: School of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea.
authors.forename: Junho
authors.initials: J
authors.lastname: Choe
authors.forename: Nara
authors.initials: N
authors.lastname: Oh
authors.forename: Sungjin
authors.initials: S
authors.lastname: Park
authors.forename: Ye Kyung
authors.initials: YK
authors.lastname: Lee
authors.forename: Ok-Kyu
authors.initials: OK
authors.lastname: Song
authors.forename: Nicolas
authors.initials: N
authors.lastname: Locker
authors.forename: Sung-Gil
authors.initials: SG
authors.lastname: Chi
authors.forename: Yoon Ki
authors.initials: YK
authors.lastname: Kim
mesh_terms: D001483:Base Sequence; D026481:Blotting, Far-Western; D002460:Cell Line; D017931:DNA Primers; D039621:Eukaryotic Initiation Factor-3 / Q000378:metabolism*; D006801:Humans; D039502:Nuclear Cap-Binding Protein Complex / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020798:Two-Hybrid System Techniques
chemical_list: D017931:DNA Primers; D039621:Eukaryotic Initiation Factor-3; D039502:Nuclear Cap-Binding Protein Complex; D012333:RNA, Messenger