Cloning and Expression of the α Subunit of Succinyl-CoA Synthetase from Dictyostelium discoideum

Antisense RNA Inhibition of the β Subunit of theDictyostelium discoideumMitochondrial Processing Peptidase Induces the Expression of Mitochondrial Proteins

“…This suggests that the -MPP synthesized and imported into the mitochondria is protected from degradation by proteases localized in the mitochondria, because the -MPP protein forms a complex with -MPP and/or is embedded in the inner membrane as part of the cytochrome bc 1 complex. In addition to Dd-MPP, similar expression patterns of mRNA and protein during development are also found in some other Dictyostelium mitochondrial proteins, such as the -subunit of succinyl-CoA synthetase, 38) DNA topoisomerase, 39) adenine nucleotide translocator, 40) and chaperonin 60, 20) exhibiting the stable nature of mitochondrial proteins in Dictyostelium development.…”

Genetic Evidence for the Expression of ATP- and GTP-specific Succinyl-CoA Synthetases in Multicellular Eucaryotes

“…The initial PCR product of 605 bp was obtained using a sense primer (5Ј-CCTATCTTTAATACTGTGAAGG-3Ј) corresponding to the peptide sequence PVFNTVKE that is found in the rat and human proteins (GenBank TM accession numbers J03621 and Z68204) and an antisense primer (5Ј-GCRTGNCCAT-3Ј) for the peptide sequence MGHA, which is conserved in procaryotes and eucaryotes and contains the active-site histidine (His-246 in E. coli) (18). The 605-bp sequence was used to design gene-specific primers for use in 3Ј-RACE; the sense primer was 5Ј-GAAATCGGAGGAAATGCTG-3Ј (corresponding to the peptide sequence EIGGNAE), which was used with a 3Ј-RACE adaptor primer (5Ј-CGTCAATGACGCATGCGACGTCCATTAG-3Ј) to obtain a 504-bp product.…”

“…The ␣-subunit is phosphorylated on a histidinyl residue as a part of the catalytic cycle (17). Recent workers have continued to assume that the ␣-subunit possesses the determinants of nucleotide specificity (18), a possibility we have shown to be untenable (15). The crystallographic structure of the E. coli enzyme is available (19).…”

Localization of a DNA topoisomerase II to mitochondria inDictyostelium discoideum: Deletion mutant analysis and mitochondrial targeting signal presequence

“…Most eukaryotic DNA topoisomerase II have a Cterminal region containing many hydrophilic and charged residues, which is thought to be a nuclear localization sequence. TopA of D. discoideum, however, lacked this C-terminal region, but had a long N-terminus containing a sequence structurally analogous to the mitochondrial targeting presequence seen in other nuclear-encoded mitochondrial proteins in D. discoideum (Troll et al 1993, Birney andKlein 1995). N-terminal deletion mutant analyses in this study also revealed that TopA is localized to the mitochondria by virtue of this additional N-terminal region (Figs.…”