OJBIPHY  Vol.11 No.4 , October 2021
The Possible Involvement of Apoptotic Decay of Terminal Deoxynucleotidyl Transferase-Positive Lymphocytes in the Reutilization of the Extracellular DNA Fragments by Surrounding Living Cells
Abstract: The migrating TdT+ thymocytes can die in other tissues, promoting the surrounding cells’ renewing likes holocrine secretion does. To clarify the role of TdT-enzyme for this function of progenitor lymphocytes, their extracellular media with its components included by living cells analyzed in vitro before and after in vivo irradiation of donor rats. The nucleoid with DNase-sensitive (free) DNA and TdT activity discovered in extracellular media conditioned preliminary by spontaneous apoptotic death of a minor part of the thymocyte’s suspension in vitro. The penetration of labeled products of non-template synthesis with free DNA’ primers from media into cells by pinocytosis confirmed by exogenous polymeric DNA marked artificially. The DNA penetration into cells follows an increase of the cell’s viability and acceleration of spontaneous intracellular DNA-synthesis controlled with labeled thymidine uptake. Both phenomena are typical for either the lowest initial concentration of intact cells or their preliminary irradiation in vivo. The data point to possible involvement of apoptotic decay of TdT+ cells in the reutilization of the extracellular DNA fragments for reparation/regeneration of surrounding living cells.
Cite this paper: Shoutko, A. (2021) The Possible Involvement of Apoptotic Decay of Terminal Deoxynucleotidyl Transferase-Positive Lymphocytes in the Reutilization of the Extracellular DNA Fragments by Surrounding Living Cells. Open Journal of Biophysics, 11, 371-382. doi: 10.4236/ojbiphy.2021.114014.

[1]   Azouna, N.B., Berraeis, L., Regaya, Z. and Jenhani, F. (2011) Immunophenotyping of Hematopoietic Progenitor Cells: Comparison between Cord Blood and Adult Mobilized Blood Grafts. World Journal of Stem Cells, 3, 104-112.

[2]   Srivastava, B.I.S. (1974) Deoxynucleotide-Polymerizing Enzymes in Normal and Malignant Human Cells. Cancer Research, 34, 1015-1026.

[3]   Haruna, T. and Atsushi, T. (2019) Age-Dependent Modification of Intracellular Zn2+ Buffering in the Hippocampus and Its Impact. Biological and Pharmaceutical Bulletin, 42, 1070-1075.

[4]   Spigelman, Z., Duff, R., Beardsley, G.P., Broder, S., Cooney, D., Landau, N.R., et al. (1988) 2’,3’-Dideoxyadenosine Is Selectively Toxic for TdT-Positive Cells. Blood, 71, 1601-1608.

[5]   Zhang, Y., Shi, M., Wen, Q., Luo, W., Yang, Z., Zhou, M., et al. (2012) Antigenic Stimulation Induces Recombination Activating Gene 1 and Terminal Deoxynucleotidyl Transferase Expression in a Murine T-Cell Hybridoma. Cellular Immunology, 274, 19-25.

[6]   McElhinny, S.A.N. and Ramsden, D.A. (2003) Polymerase Mu Is a DNA-Directed DNA/RNA Polymerase. Molecular and Cellular Biology, 23, 2309-2315.

[7]   Motea, E.A. and Berdis, A.J. (2010) Terminal Deoxynucleotidyl Transferase: The Story of a Misguided DNA Polymerase. Biochimica et Biophysica Acta (BBA)—Proteins and Proteomics, 1804, 1151-1166.

[8]   Nagler, M., Insam, H., Pietramellara, G. and Ascher-Jenull, J. (2018) Extracellular DNA in Natural Environments: Features, Relevance and Applications. Applied Microbiology and Biotechnology, 102, 6343-6356.

[9]   Zhivotovsky, B. (2020) Programmed Cell Death: Historical Notes from Russia. Biochemistry (Moscow), 85, 1127-1133.

[10]   Sheehy, E.J., Vinardell, T., Toner, M.E., Buckley, C.T. and Kelly, D.J. (2014) Altering the Architecture of Tissue Engineered Hypertrophic Cartilaginous Grafts Facilitates Vascularisation and Accelerates Mineralisation. PLoS ONE, 9, e90716.

[11]   Davila, C. and Charles, P. (1965) The Chromatography of Nucleic Acid Preparations on Deae-Cellulose Paper: I. Fractionation of Deoxyribonucleic Acid on Paper Strips or Centrifuged Paper Pulp. Journal of Chromatography A, 19, 382-395.

[12]   Doppler-Bernardi, F. and Felsenfeld, G. (1969) In Vitro Incorporation of Tritium into Native DNA. Biopolymers, 8, 733-741.

[13]   Shutko, A.N. and Shatinina, N.N. (1974) Effect of the Degree of Polymerization of Exogenous DNA on Its Incorporation into Rat Thymocytes in Vitro. Bulletin of Experimental Biology and Medicine, 77, 269-271.

[14]   Sefton, B.M. (2001) Labeling Cultured Cells with 32Pi and Preparing Cell Lysates for Immunoprecipitation. Current Protocols in Protein Science, 10, 13.2.1-13.2.8.

[15]   Shutko, A.N., Shatinina, N.N. and Rakitianskaia, I.A. (1983) Role of Terminal Deoxyri-bonucleotidyl Transferase in Stimulating Lymphocyte DNA Synthesis as Affected by PHA. Tsitologiia, 25, 1212-1215. (In Russian)

[16]   Liu, B. and Großhans, J. (2019) The Role of dNTP Metabolites in Control of the Embryonic Cell Cycle. Cell Cycle, 18, 2817-2827.

[17]   Kohnken, R., Kodigepalli, K.M. and Wu, L. (2015) Regulation of Deoxynucleotide Metabolism in Cancer: Novel Mechanisms and Therapeutic Implications. Molecular Cancer, 14, Article No. 176.

[18]   Traut, T.W. (1994) Physiological Concentrations of Purines and Pyrimidines. Molecular and Cellular Biochemistry, 140, 1-22.

[19]   Malkinson, F.D. (1981) Some Principles of Radiobiology: A Selective Review. Journal of Investigative Dermatology, 76, 32-38.

[20]   Fujita, K., Kawarada, Y., Terada, K., Sugiyama, T., Ohyama, H. and Yamada, T. (2000) Quantitative Detection of Apoptotic Thymocytes in Low-Dose X-Irradiated Mice by an Anti-Single-Stranded DNA Antibody. Journal of Radiation Research, 41, 139-149.

[21]   Ivannik, B.P., Golubeva, R.V., Proskuryakov, S.Y. and Ryabchenko, N.J. (1975) Repair and Degradation of DNA in the Irradiated Rat Thymocytes. Radiobiologiya, 15, 500-505. (Article in Russian)

[22]   Tempel, K. (1990) Changes in Nucleoid Viscosity Following X-Irradiation of Rat Thymic and Splenic Cells in Vitro. Radiation and Environmental Biophysics, 29, 19-30.

[23]   Pechatnikov, V.A., Afanasyev, V.N., Korol, B.A., Korneev, V.N., Rochev, Yu.A. and Umansky, S.R. (1986) Flow Cytometry Analysis of DNA Degradation in Thymocytes of y-Irradiated or Hydrocortisone Treated Rats. General Physiology and Biophysics, 5, 273-284.

[24]   Qiu, X., Guittet, O., Mingoes, C., El Banna, N., Huang, M.-E., Lepoivre, M. et al. (2019) Quantification of Cellular Deoxyribonucleoside Triphosphates by Rolling Circle Amplification and Förster Resonance Energy Transfer. Analytical Chemistry, 91, 14561-14568.

[25]   Huang, C.-Y., Yagüe-Capilla, M., González-Pacanowska, D. and Chang, Z.-F. (2020) Quantitation of Deoxynucleoside Triphosphates by Click Reactions. Scientific Reports, 10, Article No. 611.

[26]   Desai, A.S., Hunter, M.R. and Kapustin, A.N. (2019) Using Macropinocytosis for Intracellular Delivery of Therapeutic Nucleic Acids to Tumor Cells. Philosophical Transactions of the Royal Society B, Biological Sciences, 314, Article ID: 20180156.

[27]   Mann, C.L., Hughes, F.M. and Cidlowski, J.A. (2000) Delineation of the Signaling Pathways Involved in Glucocorticoid-Induced and Spontaneous Apoptosis of Rat Thymocytes. Endocrinology, 141, 528-538.

[28]   Shoutko, A.N., Gerasimova, O.A., Fedorov, V.A. and Zherebtsov, F.K. (2019) Non-Invasive Vibration-Stress of the Cirrhotic Liver of Patients Waiting for Transplantation Induces of Circulating CD133+ Stem Lymphocytes Committed Phenotypically toward the Liver. Open Journal of Biophysics, 9, 155-168.

[29]   Hu, Z., Chen, H., Long, Y., Li, P. and Gu, Y. (2021) The Main Sources of Circulating Cell-Free DNA: Apoptosis, Necrosis and Active Secretion. Critical Reviews in Oncology/Hematology, 157, Article ID: 103166.

[30]   Ferraro, P., Pontarin, G., Crocco, L., Fabris, S., Reichard, P. and Bianchi V. (2005) Mitochondrial Deoxynucleotide Pools in Quiescent Fibroblasts. A Possible Model for Mitochondrial Neurogastrointestinal Encephalomyopathy (MNGIE). The Journal of Biological Chemistry, 280, 24472-24480.

[31]   Deshpandea, S., Yangb, Y., Chilkotia, A. and Zauschera, S. (2019) Enzymatic Synthesis and Modification of High Molecular Weight DNA Using Terminal Deoxynucleotidyl Transferase. Methods in Enzymology, 627, 163-188.

[32]   Belli, M. and Tabocchini, M.A. (2020) Ionizing Radiation-Induced Epigenetic Modifications and Their Relevance to Radiation Protection. International Journal of Molecular Sciences, 21, Article No. 5993.

[33]   Guo, J.-R., Chen, Q.-Q., Lam, C.W.K., Wang, C.-Y., Wong, V., K.W., Chang, Z.-F., et al. (2016) Profiling Ribonucleotide and Deoxyribonucleotide Pools Perturbed by Gemcitabine in Human Non-Small Cell Lung Cancer Cells. Scientific Reports, 6, Article No. 37250.

[34]   Haghdoost, S., Czene, S., Näslund, I., Skog, S. and Harms-Ringdahl, M. (2005) Extracellular 8-oxo-dG as a Sensitive Parameter for Oxidative Stress in Vivo and in Vitro. Free Radical Research, 39, 153-162.

[35]   Sangsuwan, T. and Haghdoost, S. (2008) The Nucleotide Pool, a Target for Low-Dose γ-Ray-Induced Oxidative Stress. Radiation Research, 170, 776-783.

[36]   Cronkite, E.P., Chanana, A.D., Joel, D.D., Rai, K.R. and Schiffer, L.M. (1968) Influence of Extracorporeal Irradiation of the Blood and Lymph on Lymphopoiesis and Immunity. Proceedings of a Symposium on the Effects of Radiation on Cellular Proliferation and Differentiation, Monaco, 1-5 April 1968, 307-326.

[37]   Shoutko, A.N. (2019) Immunity or Morphogenesis in Cancer Development and Treatment. Integrative Cancer Science and Therapeutics, 6, 1-8.