A three-dimensional working model of the multienzyme complex of aminoacyl-tRNA synthetases based on electron microscopic placements of tRNA and proteins.
It has become evident that the process of protein synthesis is performed by many cellular polypeptides acting in concert within the structural confines of protein complexes. In multicellular eukaryotes, one of these assemblies is a multienzyme complex composed of eight proteins that have aminoacyl-tRNA synthetase activities as well as three non-synthetase proteins (p43, p38, and p18) with diverse functions. This study uses electron microscopy and three-dimensional reconstruction to explore the arrangement of proteins and tRNA substrates within this "core" multisynthetase complex. Binding of unfractionated tRNA establishes that these molecules are widely distributed on the exterior of the structure. Binding of gold-labeled tRNA(Leu) places leucyl-tRNA synthetase and the bifunctional glutamyl-/prolyl-tRNA synthetase at the base of this asymmetric "V"-shaped particle. A stable cell line has been produced that incorporates hexahistidine-labeled p43 into the multisynthetase complex. Using a gold-labeled nickel-nitrilotriacetic acid probe, the polypeptides of the p43 dimer have been located along one face of the particle. The results of this and previous studies are combined into an initial three-dimensional working model of the multisynthetase complex. This is the first conceptualization of how the protein constituents and tRNA substrates are arrayed within the structural confines of this multiprotein assembly.
Wolfe
Cindy L
CL
Department of Biology, Tougaloo College, Tougaloo, Mississippi 39174, USA.
Warrington
J Anthony
JA
Treadwell
Lauren
L
Norcum
Mona Trempe
MT
D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry*; D049148:Aminoacylation; D001665:Binding Sites; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000494:pharmacology; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D006639:Histidine / Q000737:chemistry; D006801:Humans; D020014:K562 Cells; D008854:Microscopy, Electron / Q000379:methods*; D008958:Models, Molecular; D009097:Multienzyme Complexes / Q000737:chemistry; D046912:Multiprotein Complexes / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D012313:RNA / Q000737:chemistry; D012343:RNA, Transfer / Q000737:chemistry
D003432:Cross-Linking Reagents; C471213:His-His-His-His-His-His; D009097:Multienzyme Complexes; D046912:Multiprotein Complexes; D009842:Oligopeptides; D010455:Peptides; D006639:Histidine; D012313:RNA; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases