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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 |
Genes addressed in this paper | ||||||||
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dut | Topics in this paper | |||||||
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 |