Sequence analysis of mitochondrial DNA in a mouse cell line resistant to chloramphenicol and oligomycin.

Abstract

A mouse L-cell line, designated 111-OB3, is described which is resistant to two drugs, chloramphenicol and oligomycin. The cells contain two types of mitochondrial DNA molecules, in roughly equal proportions, which differ in that one is cleaved by endonuclease EcoRI at a novel site within the coding sequence for subunit 6 of the mitochondrial ATPase (ATPase-6). Sequence analysis reveals that the cleavage site was created by a single transversion which predicts a replacement of valine in the wild-type ATPase-6 by glutamic acid. The replacement occurs in a hydrophobic amino acid sequence which is highly conserved in mouse, human, and bovine proteins. The position of the replacement is similar to a substitution observed in one class of yeast mutants resistant to oligomycin. Both of the mitochondrial DNA molecules in 111-OB3 also have a single nucleotide change in the gene encoding the large (16S) rRNA. These observations are consistent with the hypothesis that oligomycin resistance in mammalian cells can be cytoplasmically determined and can result from alterations in ATPase-6. The appearance of the mutation before selection in oligomycin suggests a model for the origin of mitochondrial mutations in mammalian cells.

Novel point mutations in mitochondrial 16S rRNA gene of Chinese hamster cells.

“…The four mutation sites detected in this study differ from the other six sites which have been reported in mammalian CAP-R mutants (Fig. 7) (Blanc et al, 1981a and1981b;Kearsey and Craig, 1981;Koike et al, 1983;Slott et al, 1983;Howell and Lee, 1989;Howell and Kubacka, 1993;Hashiguchi and Ikushima, 1998). It is suggested that mutation sites outside the domain may be also implicated in binding of chloramphenicol, and thus in producing the CAP-R phenotype.…”

“…(Blanc et al, 1981a and1981 b;Kearsey and Craig, 1981;Koike et al, 1983;Slott et al, 1983;Howell and Lee, 1989;Howell and Kubacka, 1993;Hashiguchi and Ikushima, 1998). To gain insight into the spontaneous mutagenesis of mammalian mtDNA, we have analyzed the spontaneous alteration of nucleotide sequences in the mitochondrial 16S rRNA genes from the CAP-R mutants isolated in Chinese hamster V79 cells without any specified mutational treatment.…”

DNA repair of UV photoproducts and mutagenesis in human mitochondrial DNA

“…In fact, there have been relatively few, if any, studies whose sole aim has been the identi®-cation of carcinogen-induced mtDNA mutations. Several studies performed in the late seventies reported the induction of speci®c mtDNA mutations in cells in culture exposed to known mutagenic agents (Spolsky & Eisenstadt, 1972;Wallace & Eisenstadt, 1979;Slott et al, 1983;Breen & Schef¯er, 1980;Blanc et al, 1981), but no information was recorded on mutation frequency or type. In 1988, Loeb and co-workers (Mita et al, 1988) attempted to measure the mutagenic potential of chemical carcinogens in mtDNA.…”

Responses of Wild-Type and Resistant Strains of the Hyperthermophilic BacteriumThermotoga maritimato Chloramphenicol Challenge

“…Note that G2288A in T. maritima corresponds to an adenine at E. coli location 2169. Mutations identified in the 23S rRNA PTC loop related to CAM resistance are shown in Table 1 for T. maritima and other organisms/organelles: G2032A in E. coli (16); G2057A in E. coli (18) and T. maritima (this work); A2058G and A2058U in Propionibacterium acnes (Pro) (54) and E. coli (16); G2061A in Rattus rattus (Rrat) (35); A2062C in Halobacterium halobium (Hha) (40); G2447A in E. coli (64), S. cerevisiae (Scer) (17), Thermus aquaticus (Taq) (50), and T. maritima (this work); A2451U in Mus musculus (Mmus) (33), E. coli (64), and T. aquaticus (50); C2542A in Homo sapiens (Hsap) (9); C2542U in H. halobium (40), Sulfolobus acidocaldarius (Sac) (1), and M. musculus (60); A2053C in S. cerevisiae (17) and E. coli (70); and U2054C in E. coli (70) and H. sapiens (9,33). elongation factor (TM1502, TM1503, and TM1590), methionine aminopeptidase (TM1478), and SecY (TM1480) genes.…”