Abstract
We have analyzed the pattern of AluI digestion over time on human chromosomes in order to monitor the evolution of the in situ enzyme action. Short treatments followed by Giemsa staining produce a G-like banding effect, whereas longer treatments produce a C-like banding pattern. However, when Propidium iodide staining is used, it reveals a uniform bright fluorescence after short AluI digestions and C bands when longer treatments are developed. We propose that C banding is the result of a uniform DNA removal in non centromeric regions taking place after a critical time point, the initial G like banding being produced by changes in the DNA-proteins interactions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Babu, A., 1988. Heterogeneity of heterochromatin of human chromosomes as demonstrated by restriction endonuclease treatment. Heterochromatin: Molecular and structural aspects. Edited by R. S. Verma, Cambridge Univ Press, pp. 250–275.
Babu, A. & Verma, R. S., 1986. The heteromorphic marker on chromosome 18 using restriction endonuclease AluI. Am. J. Hum. Genet. 38: 549–554.
Babu, A., Agarwal, A. K. & Verma, R. S., 1988. A new approach in recognition of heterochromatic regions of human chromosomes by means of restriction endonucleases. Am. J. Hum. Genet. 42: 60–65.
Babu, A., Verma, R. S. & Patil, S. R., 1987. AluI resistant chromatin on chromosome 18: classification, frequencies and implications. Chromosoma 95: 163–166.
Bianchi, N. O., Bianchi, M. S. & Cleaver, J. E., 1984. The action of ultraviolet light on the patterns of banding induced by restriction endonucleases in human chromosomes. Chromosoma 90: 133–138.
Bianchi, M. S., Bianchi, N. O., Pantelias, G. E. & Wolff, S., 1985. The mechanism and pattern of banding induced by restriction endonucleases in human chromosomes. Chromosoma 91: 131–136.
Burkholder, G. D., 1989. Morphological and biochemical effects of endonucleases on isolated mammalian chromosomes ‘in vitro’. Chromosoma 97: 347–355.
Burkholder, G. D. & Duczek, L. L., 1982. Proteins in chromosome banding I. Effect of G banding treatments on the proteins of isolated nuclei. Chromosoma 79: 29–41.
Burkholder, G. D. & Schmidt, G. J., 1986. Endonuclease banding of isolated mammalian metaphase chromosomes. Expl Cell Res. 164: 379–387.
Commings, D. D. & Avelino, E., 1974. Mechanisms of chromosome banding II. Evidence that histones are not involved. Expl Cell Res. 86: 202–206.
Cowden, R. R. & Curtis, S. K., 1981. Microfluorometric investigations of chromatin structure. I Evaluation of nine DNA specific fluorochromes as probes of chromatin organization. Histochemistry 72: 11–23.
Dyer, K. A., Riley, D. & Gartler, S. M., 1985. Analysis of inactive X chromosome structure by in situ nick translation. Chromosoma 79: 225–239.
Ferrucci, L., Romano, E. & de Stefano, G. F., 1987. AluI induced bands in great apes and man: implication for heterochromatin characterization and satellite DNA distribution. Cytogenet. Cell Genet. 44: 53–57.
Gosálvez, J., López-Fernández, C., Ferrucci, L. & Mezzanotte, R., 1989. DNA base sequence is not the only factor for restriction endonuclease activity on metaphase chromosomes: evidence using isoschizomers. Cytog. Cell Genet. 50: 142–144.
Hederman, U., Schurmann, M. & Schwinger, E., 1988. The effect of restriction enzyme digestion of human metaphase chromosomes on C-band variants of chromsomes 1 and 9. Genome 30: 652–655.
Lica, L. & Hamkalo, B., 1983. Preparation of centromeric heterochromatin by restriction endonuclease digestion of mouse L929 cells. Chromosoma 88: 42–49.
Mezzanotte, R., Bianchi, U., Vanni, R. & Ferrucci, L., 1983a. Chromatin organization and restriction endonuclease activity on human metaphase chromosomes. Cytogenet. Cell Genet. 36: 562–566.
Mezzanotte, R., Ferrucci, L., Vanni, R. & Bianchi, U., 1983b. Selective digestion of human metaphase chromosomes by Alul restriction endonuclease. J. Histochem. Cytochem. 31: 553–556.
Mezzanotte, R., Ferrucci, L., Vanni, R. & Summer, A. T., 1985. Some factors affecting the action of restriction endonucleases on human metaphase chromosomes. Expl Cell Res. 161: 247–253.
Mezzanotte, R., Peretti, D., Ennas, M. G., Vanni, R. & Summer, A. T., 1989. Analysis of human chromosome metaphase chromosomes using antibodies to double-stranded and single-stranded DNA: staining patterns are related to DNA conformation. Cytogenet. Cell Genet. 50: 54–58.
Miller, D. A., Choi, Y. & Miller, O. J., 1983. Chromosome localization of highly repetitive human DNA's & amplified ribosomal DNA with restriction enzymes. Science 219: 395–397.
Miller, D. A., Gosden, J. R., Hastie, N. D. & Evans, H. J., 1984. Mechanism of endonuclease banding of chromosomes. Expl. Cell Res. 155: 294–298.
Sumner, A. T., 1972. A simple technique for demonstrating centromeric heterochromatin. Expl. Cell Res. 75: 304–306.
Sumner, A. T., 1982. The nature and mechanisms of chromosome banding. Cancer Genet. Cytogenet. 6: 59–87.
Sumner, A. T., Evans, H. J. & Buckland, R. A., 1973. Mechanisms involved in the banding of chromosomes with Quinacrine and Giemsa. I. The effects of fixation in methanol acetic-acid. Expl Cell Res. 81: 212–222.
Zatsepina, O. V., Polyakov, V. Y. & Chentsov, Y. S., 1989. Differential decondensation of mitotic chromosomes during hypotonic treatment of living cells as a possible cause of G-banding: an ultrastructural study. Chromosoma 98: 109–116.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fernández-Peralta, A.M., Tagarro, I., Ludeña Reyes, P. et al. Time-dependent AluI action on human chromosomes. Genetica 84, 191–194 (1991). https://doi.org/10.1007/BF00127246
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00127246