title: Mammalian mitochondrial initiation factor 2 supports yeast mitochondrial translation without formylated initiator tRNA.
abstract: Initiation of protein synthesis in mitochondria and chloroplasts is widely believed to require a formylated initiator methionyl-tRNA (fMet-tRNAfMet) in a process involving initiation factor 2 (IF2). However, yeast strains disrupted at the FMT1 locus, encoding mitochondrial methionyl-tRNA formyltransferase, lack detectable fMet-tRNAfMet but exhibit normal mitochondrial function as evidenced by normal growth on non-fermentable carbon sources. Here we show that mitochondrial translation products in Saccharomyces cerevisiae were synthesized in the absence of formylated initiator tRNA. ifm1 mutants, lacking the mitochondrial initiation factor 2 (mIF2), are unable to respire, indicative of defective mitochondrial protein synthesis, but their respiratory defect could be complemented by plasmid-borne copies of either the yeast IFM1 gene or a cDNA encoding bovine mIF2. Moreover, the bovine mIF2 sustained normal respiration in ifm1 fmt1 double mutants. Bovine mIF2 supported the same pattern of mitochondrial translation products as yeast mIF2, and the pattern did not change in cells lacking formylated Met-tRNAfMet. Mutant yeast lacking any mIF2 retained the ability to synthesize low levels of a subset of mitochondrially encoded proteins. The ifm1 null mutant was used to analyze the domain structure of yeast mIF2. Contrary to a previous report, the C terminus of yeast mIF2 is required for its function in vivo, whereas the N-terminal domain could be deleted. Our results indicate that formylation of initiator methionyl-tRNA is not required for mitochondrial protein synthesis. The ability of bovine mIF2 to support mitochondrial translation in the yeast fmt1 mutant suggests that this phenomenon may extend to mammalian mitochondria as well.
authors.affiliation: Department of Chemistry and Biochemistry, Institute for Cellular and Molecular Biology, The University of Texas, Austin, Texas 78712, USA.
authors.forename: Anne S
authors.initials: AS
authors.lastname: Tibbetts
authors.forename: Lena
authors.initials: L
authors.lastname: Oesterlin
authors.forename: Sherwin Y
authors.initials: SY
authors.lastname: Chan
authors.forename: Gisela
authors.initials: G
authors.lastname: Kramer
authors.forename: Boyd
authors.initials: B
authors.lastname: Hardesty
authors.forename: Dean R
authors.initials: DR
authors.lastname: Appling
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D005816:Genetic Complementation Test; D008928:Mitochondria / Q000378:metabolism*; D039881:Prokaryotic Initiation Factor-2 / Q000235:genetics / Q000502:physiology*; D012331:RNA, Fungal / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D039881:Prokaryotic Initiation Factor-2; D012331:RNA, Fungal; D012343:RNA, Transfer