The interaction of mammalian mitochondrial translational initiation factor 3 with ribosomes: evolution of terminal extensions in IF3mt.
Mammalian mitochondrial initiation factor 3 (IF3(mt)) has a central region with homology to bacterial IF3. This homology region is preceded by an N-terminal extension and followed by a C-terminal extension. The role of these extensions on the binding of IF3(mt) to mitochondrial small ribosomal subunits (28S) was studied using derivatives in which the extensions had been deleted. The K(d) for the binding of IF3(mt) to 28S subunits is approximately 30 nM. Removal of either the N- or C-terminal extension has almost no effect on this value. IF3(mt) has very weak interactions with the large subunit of the mitochondrial ribosome (39S) (K(d) = 1.5 muM). However, deletion of the extensions results in derivatives with significant affinity for 39S subunits (K(d) = 0.12-0.25 muM). IF3(mt) does not bind 55S monosomes, while the deletion derivative binds slightly to these particles. IF3(mt) is very effective in dissociating 55S ribosomes. Removal of the N-terminal extension has little effect on this activity. However, removal of the C-terminal extension leads to a complex dissociation pattern due to the high affinity of this derivative for 39S subunits. These data suggest that the extensions have evolved to ensure the proper dissociation of IF3(mt) from the 28S subunits upon 39S subunit joining.
Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC-27599-3290, USA.
Md Emdadul
ME
Haque
Domenick
D
Grasso
Linda L
LL
Spremulli
D000818:Animals; D002417:Cattle; D039642:Eukaryotic Initiation Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019143:Evolution, Molecular; D006160:Guanosine Triphosphate / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008928:Mitochondria / Q000235:genetics; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D039665:Prokaryotic Initiation Factor-3 / Q000737:chemistry; D011485:Protein Binding; D012358:RNA, Transfer, Met / Q000378:metabolism; D054683:Ribosome Subunits, Large, Eukaryotic / Q000378:metabolism*; D054682:Ribosome Subunits, Small, Eukaryotic / Q000378:metabolism*; D017384:Sequence Deletion; D020349:Surface Plasmon Resonance
D039642:Eukaryotic Initiation Factors; C503923:GDPNP compound; C519156:MTIF3 protein, human; D024101:Mitochondrial Proteins; D039665:Prokaryotic Initiation Factor-3; D012358:RNA, Transfer, Met; D006160:Guanosine Triphosphate