Reversible permeabilization of lymphocytes destroys the incorporation of deoxythymidine but not of deoxycytidine

J. Taljanidisz, M. Sasvári, Tatjana Spasokutotskaja, F. Antoni, M. Staub

Research output: Contribution to journalArticle

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Abstract

The total uptake, phosphorylation and incorporation of thymidine (dThd) and deoxycytidine (dCyd) were compared in intact and reversibly permeabilized human tonsillar lymphocytes. The total uptake of [3H]dThd was lower than that of [5-3H]dCyd, but almost all of [3H]dThd was incorporated into DNA. However, the main part of [5-3H]dCyd taken up by the lymphocytes was found in the pool as phosphorylated nucleoside (55%), and only a smaller part (13%) was incorporated into DNA. Phosphorylated nucleosides were determined by DEAE-cellulose sheets in the ethanol-soluble fraction of the cells. The reversible permeabilization of lymphocytes by Dextran T-150 destroys totally the [3H]dThd incorporation, while [5-3H]dCyd incorporation decreased only to 60% of intact cells. During permeabilization the phosphorylation of both nucleosides increased severalfold. After permeabilization all [3H]dThd was in dTMP form, while [5-3H]dCyd was also found in dCDP (3%) and dCTP (38%) form. In the meanwhile, 22% of thymidine kinase, 63% of deoxycytidine kinase and 98% of DNA polymerase activity were measured in permeabilized cells as compared to intact cells. The results suggest different relationships between the lymphocyte plasma membrane and the salvage pathways of the two pyrimidine nucleosides.

Original languageEnglish
Pages (from-to)266-271
Number of pages6
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume885
Issue number3
DOIs
Publication statusPublished - Mar 14 1986

Fingerprint

Deoxycytidine
Thymidine
Lymphocytes
Nucleosides
Deoxycytidine Kinase
Phosphorylation
Pyrimidine Nucleosides
DEAE-Cellulose
Thymidine Kinase
DNA
DNA-Directed DNA Polymerase
Dextrans
Ethanol
Cell Membrane

Keywords

  • (Lymphocyte)
  • Deoxycytidine
  • Deoxythymidine
  • Permeabilization
  • Phosphorylation

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Reversible permeabilization of lymphocytes destroys the incorporation of deoxythymidine but not of deoxycytidine. / Taljanidisz, J.; Sasvári, M.; Spasokutotskaja, Tatjana; Antoni, F.; Staub, M.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 885, No. 3, 14.03.1986, p. 266-271.

Research output: Contribution to journalArticle

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AB - The total uptake, phosphorylation and incorporation of thymidine (dThd) and deoxycytidine (dCyd) were compared in intact and reversibly permeabilized human tonsillar lymphocytes. The total uptake of [3H]dThd was lower than that of [5-3H]dCyd, but almost all of [3H]dThd was incorporated into DNA. However, the main part of [5-3H]dCyd taken up by the lymphocytes was found in the pool as phosphorylated nucleoside (55%), and only a smaller part (13%) was incorporated into DNA. Phosphorylated nucleosides were determined by DEAE-cellulose sheets in the ethanol-soluble fraction of the cells. The reversible permeabilization of lymphocytes by Dextran T-150 destroys totally the [3H]dThd incorporation, while [5-3H]dCyd incorporation decreased only to 60% of intact cells. During permeabilization the phosphorylation of both nucleosides increased severalfold. After permeabilization all [3H]dThd was in dTMP form, while [5-3H]dCyd was also found in dCDP (3%) and dCTP (38%) form. In the meanwhile, 22% of thymidine kinase, 63% of deoxycytidine kinase and 98% of DNA polymerase activity were measured in permeabilized cells as compared to intact cells. The results suggest different relationships between the lymphocyte plasma membrane and the salvage pathways of the two pyrimidine nucleosides.

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