Kinectin-dependent assembly of translation elongation factor-1 complex on endoplasmic reticulum regulates protein synthesis.
Kinectin is an integral membrane protein with many isoforms primarily found on the endoplasmic reticulum. It has been found to bind kinesin, Rho GTPase, and translation elongation factor-1delta. None of the existing models for the quaternary organization of the elongation factor-1 complex in higher eukaryotes involves kinectin. We have investigated here the assembly of the elongation factor-1 complex onto endoplasmic reticulum via kinectin using in vitro and in vivo assays. We established that the entire elongation factor-1 complex can be anchored to endoplasmic reticulum via kinectin, and the interacting partners are as follows. Kinectin binds EF-1delta, which in turn binds EF-1gamma but not EF-1beta; EF-1gamma binds EF-1delta and EF-1beta but not kinectin. In vivo splice blocking of the kinectin exons 36 and 37 produced kinectin lacking the EF-1delta binding domain, which disrupted the membrane localization of EF-1delta, EF-1gamma, and EF-1beta on endoplasmic reticulum, similar to the disruptions seen with the overexpression of kinectin fragments containing the EF-1delta binding domain. The disruptions of the EF-1delta/kinectin interaction inhibited expression of membrane proteins but enhanced synthesis of cytosolic proteins in vivo. These findings suggest that anchoring the elongation factor-1 complex onto endoplasmic reticulum via EF-1delta/kinectin interaction is important for regulating protein synthesis in eukaryotic cells.
Ong
Lee-Lee
LL
National University Medical Institutes, National University of Singapore, Singapore 117597, Singapore.
Lin
Pao-Chun
PC
Zhang
Xin
X
Chia
Ser-Mien
SM
Yu
Hanry
H
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis / Q000235:genetics*; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
C088547:KTN1 protein, human; D008565:Membrane Proteins; D020648:Peptide Elongation Factor 1; D021122:Protein Subunits; D012333:RNA, Messenger