Induction of Chromosomal Aberrations by the Restriction Endonuclease AluI in Chinese Hamster Ovary (CHO) Cells: Influence of Glycerol on Aberration Frequencies

  • C. Johannes
  • G. Obe


Restriction endonucleases (REs) induce chromosomal aberrations in living cells indicating that the ultimate lesion for the production of chromosomal aberrations is a DNA double-strand break (Obe et al. 1987; Bryant, this Vol.; Morgan et al., this Vol.). It is still not clear how living cells can uptake REs. Different methods have been used to treat cells with REs (for discussion see Bryant 1988; Morgan et al., this Vol.) and all of them are more or less successful when measured as frequencies of chromosomal aberrations induced by REs. From our own results concerning the induction of chromosomal aberrations with AluI following treatment of pellets of 4 × 106 cells (about 10 µl volume) with 8 µl of a suspension of REs and newborn calf serum, we proposed that REs are internalized via an energy-dependent receptor-mediated mechanism (Johannes et al. 1989; Obe et al. 1989; Vasudev and Obe 1987). Analyses of Bryant and Christie (1989) have shown that the uptake of REs using the “pellet technique” is dependent on the shipping buffer in which the REs are suspended. The authors assume that “this may result from a combination of hypertonic shock and the detergent present in the shipping buffer” (Bryant and Christie 1989). In order to avoid a possible influence on the uptake of AluI by the relatively high concentrations of buffer components we analyzed the chromosome breaking effects of AluI by treatment of monolayers in such a way that the concentrations of the buffer components are considerably lower when compared to our earlier method.


Chromosomal Aberration Chinese Hamster Ovary Cell Chinese Hamster Ovary Newborn Calf Serum Dicentric Chromosome 
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  1. Bryant PE (1988) Use of restriction endonucleases to study relationships between DNA double-strand breaks, chromosomal aberrations and other end-points in mammalian cells. Int J Radiat Biol 54: 869–890PubMedCrossRefGoogle Scholar
  2. Bryant PE, Christie AF (1989) Induction of chromosomal aberrations in CHO cells by restriction endonucleases: effects of blunt- und cohesive-ended double-strand breaks in cells treated by “pellet” method. Mutat Res 213: 233–241PubMedCrossRefGoogle Scholar
  3. Fraley R, Straubinger RM, Rule G, Springer EL Papahadjopoulos D (1981) Liposome-mediated delivery of deoxyribonucleic acid to cells: enhanced efficiency of delivery related to lipid composition and incubation conditions. Biochemistry 20: 6978–6987PubMedCrossRefGoogle Scholar
  4. Galloway SM, Deasy DA, Bean CL, Kraynak AR, Armstrong MJ, Bradley MO (1987) Effects of high osmotic strength on chromosome aberrations, sister-chromatid exchanges and DNA strand breaks, and the relation to toxicity. Mutat Res 189: 15–25PubMedCrossRefGoogle Scholar
  5. Johannes C, Tuschy S, Lamprecht I, Obe G (1989) The intercellular distributions of chromosomal aberrations induced by the restriction endonucleases AluI and Bam HI in Chinese hamster ovary cells are overdispersed. Biol Zentralbl 108: 445–451Google Scholar
  6. Obe G, Vasudev V, Johannes C (1987) Chromosome aberrations induced by restriction endonucleases. In: Obe G, Basler A (eds) Cytogenetics, basic and applied aspects. Springer, Berlin Heidelberg New York, pp 300–314Google Scholar
  7. Obe G, Besoke R, Schmidt S (1989) Receptor-mediated uptake of AluI by Chinese hamster ovary cells: evidence from experiments with various treatment times and treatment temperatures, and with chloroquine, methylamine and monensin. Biol Zentralbl 108: 439–444Google Scholar
  8. Vasudev V, Obe G (1987) Evidence for a receptor-mediated endocytosis of AluI in Chinese hamsterovary cells. Mutat Res 197: 109–116Google Scholar
  9. Winegar RA, Philips JW, Youngblom JH, Morgan W (1989) Cell electroporation is a highly efficient method for introducing restriction endonucleases into cells. Mutat Res 225: 49–53PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1990

Authors and Affiliations

  • C. Johannes
  • G. Obe
    • 1
  1. 1.Department of GeneticsUniversity-GH EssenEssen 1Germany

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