TY - JOUR
T1 - Interaction of mitochondrial initiation factor 2 with mitochondrial fMet-tRNA
AU - Spencer, Angela C.
AU - Spremulli, Linda L.
PY - 2004
Y1 - 2004
N2 - The mammalian mitochondrial genome contains a single tRNAMet gene Nat gives rise to the initiator and elongator tRNAMet. It is generally believed that mitochondrial protein synthesis begins with formyl-methionyl-tRNA, which indicates that the formylation of mitochondrial Met-tRNA specifies its participation in initiation through its interaction with initiation factor 2 (IF-2). However, recent studies in yeast mitochondria, suggest that formylation is not required for protein synthesis. In addition, bovine IF-2mt could replace yeast IF-2mt in strains that lack fMet-tRNA which suggests that this paradigm may extend to mammalian mitochondria. Here, the importance of the formylation of mitochondrial Met-tRNA for the interaction with IF-2mt was investigated by measuring the ability of bovine IF-2mt to bind mitochondrial fMet-tRNA. In direct binding experiments, bovine IF-2mt has a 25-fold greater affinity for mitochondrial fMet-tRNA than Met-tRNA, using either the native mitochondrial tRNAMet or an in vitro transcript of bovine mitochondrial tRNAMet. In addition, IF-2mt will not effectively stimulate mitochondrial Met-tRNA binding to mitochondrial ribosomes, exhibiting a 50-fold preference for fMet-tRNA over Met-tRNA in this assay. Finally, the region of IF-2mt responsible for the interaction with fMet-tRNA was mapped to the C2 sub-domain of domain VI of this factor.
AB - The mammalian mitochondrial genome contains a single tRNAMet gene Nat gives rise to the initiator and elongator tRNAMet. It is generally believed that mitochondrial protein synthesis begins with formyl-methionyl-tRNA, which indicates that the formylation of mitochondrial Met-tRNA specifies its participation in initiation through its interaction with initiation factor 2 (IF-2). However, recent studies in yeast mitochondria, suggest that formylation is not required for protein synthesis. In addition, bovine IF-2mt could replace yeast IF-2mt in strains that lack fMet-tRNA which suggests that this paradigm may extend to mammalian mitochondria. Here, the importance of the formylation of mitochondrial Met-tRNA for the interaction with IF-2mt was investigated by measuring the ability of bovine IF-2mt to bind mitochondrial fMet-tRNA. In direct binding experiments, bovine IF-2mt has a 25-fold greater affinity for mitochondrial fMet-tRNA than Met-tRNA, using either the native mitochondrial tRNAMet or an in vitro transcript of bovine mitochondrial tRNAMet. In addition, IF-2mt will not effectively stimulate mitochondrial Met-tRNA binding to mitochondrial ribosomes, exhibiting a 50-fold preference for fMet-tRNA over Met-tRNA in this assay. Finally, the region of IF-2mt responsible for the interaction with fMet-tRNA was mapped to the C2 sub-domain of domain VI of this factor.
UR - http://www.scopus.com/inward/record.url?scp=6044234880&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=6044234880&partnerID=8YFLogxK
U2 - 10.1093/nar/gkh886
DO - 10.1093/nar/gkh886
M3 - Article
C2 - 15477394
AN - SCOPUS:6044234880
SN - 0305-1048
VL - 32
SP - 5464
EP - 5470
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 18
ER -