Cell cycle- and differentiation stage-dependent variation of dUTPase activity in higher plant cells

Abstract

Deoxyuridine triphosphate nucleotidohydrolase (dUTPase), a key enzyme in pyrimidine nucleotide metabolism, specifically hydrolyzes deoxyuridine triphosphate (dUTP) to deoxyuridine monophosphate and inorganic pyrophosphate. This enzyme activity has been studied in cellular extracts from Allium cepa root meristem cells with two specific aims: (i) to determine how the properties of the plant enzyme compare with those of dUTPase purified from other sources, and (ii) to analyze the relationship between dUTPase activity and cell proliferation and cell differentiation. Plant dUTPase is highly specific for dUTP, with an apparent Km of 6 microM, is highly sensitive to EDTA and it is probably a metalloenzyme. Our results demonstrate the presence of high levels of dUTPase in both resting and proliferating root meristem cells. The enzyme activity appears to be tightly regulated during the cell cycle. dUTPase activity increases at the G1/S boundary, remains high throughout S phase, and shows almost undetectable levels during G1 and G2. We have also found that dUTPase activity in differentiated cells, located in the mature portion of the root, is barely detectable. Altogether our results indicate that dUTPase activity is modulated by the proliferation rate and that this activity progressively decreases as cells initiate their differentiation program.