Nascent DNA chains synthesized in reversibly permeable cells of mouse thymocytes.

G. Bánfalvi, A. Sooki-Toth, N. Sarkar, S. Csuzi, F. Antoni

Research output: Contribution to journalArticle

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Abstract

Freshly prepared thymocytes continue to synthesize DNA under hypotonic conditions in the presence of 4.5% dextran T-150, the four deoxyribonucleoside triphosphates and ATP. Permeable cells could seal the membrane in a serum-enriched medium within a few hours. 2'-Deoxycytidine 5'-triphosphate is effectively substituted by 5-mercuri-2'-deoxycytidine 5'-triphosphate as a substrate. The newly synthesized mercurated DNA can be separated from cellular DNA and RNA on a thiol-agarose affinity matrix. The rate of incorporation of [3H]thymidine triphosphate into permeable cells is the same as that of the incorporation of [3H]thymidine into intact cells, corresponding to approximately 30% of the rate in vivo. Synthesis in permeable cells reflects DNA replication shown by inhibitors such as 1-beta-D-arabinofuranosylcytosine 5'-triphosphate (aCTP), nalidixic acid and novobiocin and by density shift experiments. More than 80% of the newly synthesized low-molecular-mass DNA, 8-60 nucleotides in length, consists of RNA-linked DNA. This conclusion is based on phosphorylation with [gamma-32]ATP and polynucleotide kinase and rephosphorylation after alkaline hydrolysis. The 5' end of RNA consists of adenylate, guanylate, cytidylate and uridylate residues in a ratio of 4:3:1.5:1.5.

Original languageEnglish
Pages (from-to)553-559
Number of pages7
JournalEuropean Journal of Biochemistry
Volume139
Issue number3
Publication statusPublished - Mar 15 1984

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Thymocytes
Cells
DNA
RNA
Arabinofuranosylcytosine Triphosphate
Adenosine Triphosphate
Polynucleotide 5'-Hydroxyl-Kinase
Deoxyribonucleosides
Novobiocin
Nalidixic Acid
Dextrans
Phosphorylation
DNA Replication
Sulfhydryl Compounds
Sepharose
Thymidine
Cytarabine
Molecular mass
Hydrolysis
Nucleotides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nascent DNA chains synthesized in reversibly permeable cells of mouse thymocytes. / Bánfalvi, G.; Sooki-Toth, A.; Sarkar, N.; Csuzi, S.; Antoni, F.

In: European Journal of Biochemistry, Vol. 139, No. 3, 15.03.1984, p. 553-559.

Research output: Contribution to journalArticle

Bánfalvi, G, Sooki-Toth, A, Sarkar, N, Csuzi, S & Antoni, F 1984, 'Nascent DNA chains synthesized in reversibly permeable cells of mouse thymocytes.', European Journal of Biochemistry, vol. 139, no. 3, pp. 553-559.
Bánfalvi, G. ; Sooki-Toth, A. ; Sarkar, N. ; Csuzi, S. ; Antoni, F. / Nascent DNA chains synthesized in reversibly permeable cells of mouse thymocytes. In: European Journal of Biochemistry. 1984 ; Vol. 139, No. 3. pp. 553-559.
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