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Nuclear inheritance of oligomycin resistance in mouse L cells
Abstract:
The inheritance of oligomycin resistance was studied in three mouse L-cell mutants, OLI 2, OLI 4, and OLI 14. All three mutants had previously been shown to have oligomycin-resistant mitochondrial ATPase activity. In addition, OLI 14 has DCCD-resistant mitochondrial ATPase activity and an altered DCCD-binding protein. Oligomycin-resistant cells were enucleated and fused with oligomycin-sensitive cells under a variety of selective regimes designed to allow growth of oligomycin-resistant cybrids. No transfer of oligomycin resistance via the cytoplasm of OLI 2, OLI 4, or OLI 14 was detected. In contrast, oligomycin resistance was transferred with the karyoplasts of OLI 14 in karyoplast-cell fusions. Fusions between OLI 14 cells and oligomycin-sensitive cells also produced oligomycin-resistant hybrids. Transfer of oligomycin resistance in the karyoplast-cell and cell-cell fusions were demonstrated at the level of the mitochondrial ATPase. These results indicate that oligomycin resistance in OLI 14 is most likely under nuclear control. Furthermore, nuclear inheritance of oligomycin resistance in a mutant with a modified DCCD-binding protein suggests that the gene for the DCCD-binding protein is encoded in the nucleus of mammalian cells.
Insights
Oligomycin resistance in mouse cells was investigated. Results show resistance is inherited through the nucleus, not cytoplasm, suggesting nuclear control of mitochondrial function and DCCD-binding protein genes.
Area of Science:
- Cell Biology
- Genetics
- Biochemistry
Background:
- Mitochondrial ATPase activity can confer resistance to oligomycin.
- Mouse L-cell mutants OLI 2, OLI 4, and OLI 14 exhibit oligomycin-resistant mitochondrial ATPase activity.
- OLI 14 mutant displays additional resistance to DCCD and an altered DCCD-binding protein.
Purpose of the Study:
- To investigate the inheritance pattern of oligomycin resistance in mouse L-cell mutants.
- To determine whether oligomycin resistance is maternally (cytoplasmic) or paternally (nuclear) inherited.
- To elucidate the genetic control of the DCCD-binding protein.
Main Methods:
- Cell fusion experiments involving enucleated oligomycin-resistant cells and intact sensitive cells.
- Karyoplast-cell fusions using OLI 14 karyoplasts and sensitive cells.
- Cell-cell fusions between OLI 14 and sensitive cells.
- Analysis of mitochondrial ATPase activity to confirm resistance transfer.
Main Results:
- No transfer of oligomycin resistance was observed through the cytoplasm of OLI 2, OLI 4, or OLI 14 cells.
- Oligomycin resistance was successfully transferred via karyoplasts of OLI 14 in karyoplast-cell fusions.
- Oligomycin-resistant hybrids were generated from fusions between OLI 14 and sensitive cells.
- Transfer of resistance was confirmed at the mitochondrial ATPase level.
Conclusions:
- Oligomycin resistance in the OLI 14 mutant is primarily under nuclear control.
- The gene encoding the DCCD-binding protein is likely located in the nucleus of mammalian cells.
- This suggests nuclear inheritance of mitochondrial traits in mammals.