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Chia, CP Catherine P, Inoguchi, Noriko, Varon, KC Kyle C, Bartholomai, BM Bradley M, Moriyama, Hideaki, (2020) ' Mitochondrial localization of Dictyostelium discoideum dUTPase mediated by its N-terminus. ' BMC Res Notes 13 16
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Abstract:The annotated genome of D. discoideum identifies a gene encoding a polypeptide containing the five conserved motifs of homotrimeric dUTPases. Recombinant proteins, comprised of either full-length or core polypeptides with all conserved motifs but lacking residues 1-37 of the N-terminus, were active dUTPases. Crystallographic analyses of the core enzyme indicated that the C-termini, normally flexible, were constrained by interactions with the shortened N-termini that arose from the loss of residues 1-37. This allowed greater access of dUTP to active sites, resulting in enhanced catalytic parameters. A tagged protein comprised of the N-terminal forty amino acids of dUTPase fused to green fluorescent protein (GFP) was expressed in D. discoideum cells. Supporting a prediction of mitochondrial targeting information within the N-terminus, localization and subcellular fractionation studies showed GFP to be in mitochondria. N-terminal sequencing of immunoprecipitated GFP revealed the loss of the dUTPase sequence upon import into the organelle.
Status: epublish Type: Journal article Source: PUBMED PubMed ID: 31910901

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Protein Physical Properties X
Mammalian Gene Related X
Protein Functional Domain X
Non-mammalian Gene Related X
Cellular Location X
Protein/Nucleic Acid Structure X
Substrates/Ligands/Cofactors X
Function/Process X