Maturation stage and proliferation-dependent expression of dUTPase in human T cells.

Strahler, John R.; Zhu, Xiaoxiang; Hora, N.A.; Wang, Y. K.; Andrews, P.; Roseman, N A; Neel, James V.; Turka, Laurence A.; Hanash, Samir M.

doi:10.1073/pnas.90.11.4991

Cited by 46 publications

(48 citation statements)

References 32 publications

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“…dUTPase in mitochondria is necessary for integrity of mitochondrial DNA that replicates independently from the cell cycle. In mature lymphocytes, correlation between dUTPase presence and cell mitogenic status suggested enzyme up-regulation in stages associated with DNA synthesis, whereas in immature populations, enzyme levels are constitutive (3). The requirement of actively dividing cells for high dUTPase levels is also confirmed by studies in rat liver regeneration (4) and mitogen-activated T and B cells (5).…”

supporting

confidence: 52%

“…However, several fractions of S2 cell extract modulated the activity of recombinant Drosophila dUTPase strictly, depending on the presence of the unique C terminus. 3 This unique region may therefore mediate regulation of dUTPase activity by other protein factors. dUTPases from various organisms contain species-specific, flexible N-or C-terminal extensions that provide interaction surface for cellular macromolecules (17)(18)(19).…”

Section: Discussionmentioning

confidence: 99%

“…Experiments performed in our laboratory also indicated that several fractions of S2 cell extract have significant modulatory effect on the recombinant 23-kDa dUTPase isoform. 3 These observations initiated investigations by two experimental approaches toward endogenous macromolecules capable of binding to dUTPase. Fig.…”

Section: Subcellular Localization Of Dutpase In Different Tissues Ofmentioning

confidence: 99%

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Developmental Regulation of dUTPase in Drosophila melanogaster

Békési

Zagyva

Hunyadi‐Gulyás

et al. 2004

Journal of Biological Chemistry

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dUTPase prevents uracil incorporation into DNA by strict regulation of the cellular dUTP:dTTP ratio. Lack of the enzyme initiates thymineless cell death, prompting studies on enzyme regulation. We investigated expression pattern and localization of Drosophila dUTPase. Similarly to human, two isoforms of the fly enzyme were identified at both mRNA and protein levels. During larval stages, a drastic decrease of dUTPase expression was demonstrated at the protein level. In contrast, dUTPase mRNAs display constitutive character throughout development. A putative nuclear localization signal was identified in one of the two isoforms. However, immunohistochemistry of ovaries and embryos did not show a clear correlation between the presence of this signal and subcellular localization of the protein, suggesting that the latter may be perturbed by additional factors. Results are in agreement with a multilevel regulation of dUTPase in the Drosophila proteome, possibly involving several interacting protein partners of the enzyme. Using independent approaches, the existence of such macromolecular partners was verified.Faithful conservation and transmission of genetic information are crucial for living organisms. The enzyme dUTPase prevents uracil incorporation into DNA by hydrolysis of dUTP into dUMP and inorganic pyrophosphate and provides a unique and essential preventive DNA repair function. Lack of dUTPase leads to uracil-substituted DNA that perturbs base excision repair, resulting in DNA fragmentation and thymineless apoptosis of the cell. The physiological role of the enzyme argues for regulation of dUTPase presence, localization, and/or function, depending on cell status.This implication was closely investigated in human cells.

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supporting

confidence: 52%

Section: Discussionmentioning

confidence: 99%

Section: Subcellular Localization Of Dutpase In Different Tissues Ofmentioning

confidence: 99%

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Developmental Regulation of dUTPase in Drosophila melanogaster

Békési

Zagyva

Hunyadi‐Gulyás

et al. 2004

Journal of Biological Chemistry

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“…More recently, Strahler and co-workers demonstrated that, upon peripheral blood lymphocyte stimulation, there is a large induction of the phosphorylated form of dUTPase. This induction of dUTPase protein coincided with the onset of DNA replication, suggesting a link between dUTPase phosphorylation and the proliferation status of the cell (4).…”

mentioning

confidence: 99%

Identification of a Consensus Cyclin-dependent Kinase Phosphorylation Site Unique to the Nuclear Form of Human Deoxyuridine Triphosphate Nucleotidohydrolase

Ladner

Carr

Huddleston

et al. 1996

Journal of Biological Chemistry

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, we identified two distinct isoforms of dUTPase in human cells. These isoforms are individually targeted to the nucleus (DUT-N) and mitochondria (DUT-M). The proteins are nearly identical, differing only in a short region of their amino termini. Despite the structural differences between these proteins, they retain identical affinities for dUTP (preceding article). In previous work, this laboratory demonstrated that dUTPase is posttranslationally phosphorylated on serine residue(s) (Lirette, R., and Caradonna, S. (1990) J. Cell. Biochem. 43, 339 -353). To extend this work and determine if both isoforms of dUTPase are phosphorylated, a more in depth analysis of dUTPase phosphorylation was undertaken. [ 32 P]Orthophosphatelabeled dUTPase was purified from HeLa cells, revealing that only the nuclear form of dUTPase is phosphorylated. Electrospray tandem mass spectrometry was used to identify the phosphorylation site as Ser-11 in the amino-terminal tryptic peptide PCSEETPAIpSPSKR (the NH 2 -terminal Met is removed in the mature protein). Mutation of Ser-11 by replacement with Ala blocks phosphorylation of dUTPase in vivo. Analysis of the wild type and Ser-11 3 Ala mutant indicates that phosphorylation does not regulate the enzymatic activity of the DUT-N protein in vitro. Additionally, experiments with the Ser-11 3 Ala mutant indicate that phosphorylation does not appear to play a role in subunit association of the nuclear form of dUTPase. The amino acid context of this phosphorylation site corresponds to the consensus target sequence for the cyclin-dependent protein kinase p34 cdc2 . Recombinant DUT-N was specifically phosphorylated on Ser-11 in vitro with immunoprecipitated p34 cdc2 . Together, these data suggest that the nuclear form of dUTPase may be a target for cyclin-dependent kinase phosphorylation in vivo.Human dUTPase 1 was first purified from HeLa cells by Caradonna and Adamkiewicz (2). The enzyme was characterized as a homodimer with a monomeric molecular weight of 22,500 and a K m for dUTP of 2.5 M. The dUTPase monomers associate in the presence of divalent cations such as magnesium or manganese to form the active enzyme. In later work, we identified the human enzyme as a serine phosphoprotein (1). Studies on herpesvirus infection of HeLa cells have shown that cellular dUTPase activity decreases postinfection, while the virus-encoded dUTPase activity increases. It was postulated that the associated decrease of cellular dUTPase activity was not due to rapid degradation but rather correlated with dephosphorylation of the host dUTPase protein. These data suggest that phosphorylation may play a role in regulating the enzymatic activity of the human dUTPase protein. More recently, Strahler and co-workers demonstrated that, upon peripheral blood lymphocyte stimulation, there is a large induction of the phosphorylated form of dUTPase. This induction of dUTPase protein coincided with the onset of DNA replication, suggesting a link between dUTPase phosphorylation and the proliferation status of the cell (4).In ...

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“…In an attempt to understand and explain our results, we found in previously published work that the expression of cellular dUTPase is regulated by the cell cycle and is at higher levels in dividing cells than in non-dividing cells (Pardo & Gutierrez, 1990;Strahler et al, 1993). In the context of virus infection, uracil incorporation controlled by the expression of cellular dUTPase and enzymes related to dTTP biosynthesis could work as a weapon against viruses (Priet et al, 2006).…”

Section: Discussionmentioning

confidence: 89%

Genômica, evolução e caracterização funcional de genes de baculovírus

Araújo¹

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Maturation stage and proliferation-dependent expression of dUTPase in human T cells.

Cited by 46 publications

References 32 publications

Developmental Regulation of dUTPase in Drosophila melanogaster

Developmental Regulation of dUTPase in Drosophila melanogaster

Identification of a Consensus Cyclin-dependent Kinase Phosphorylation Site Unique to the Nuclear Form of Human Deoxyuridine Triphosphate Nucleotidohydrolase

Genômica, evolução e caracterização funcional de genes de baculovírus

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