Abstract
Measurements of the integrated absorbancy of naphthol yellow S binding to protein (430 nm) and Feulgen-stained DNA (550 nm) of two puff regions in Drosophila hydei polytene chromosomes revealed a significant increase in the naphthol yellow S binding capacity during the first 5 min of puff induction. The ratio of integrated absorption values at 430 and 550 nm of two chromosome regions, 2-48 C and 4-81 B were determined relative to the ratio of absorption values at 430 and 550 nm of a reference band. These determinations were carried out in a non-puffed state and at 5, 10, 30, 60 and 120 min after onset of a temperature treatment inducing puffs in these regions. The quotient of the absorption ratio of the puff region and the ratio of the reference band provides a relative measure for naphthol yellow S binding to protein. The staining reaction was absent after pronase treatment.—The relative increase in naphthol yellow S binding was most obvious during the first 5 min after onset of puff induction. The binding of naphthol yellow S was increased by a factor 1.7 for puff 2-48 C, and a factor 1.9 for puff 4-81 B. The maximum value, indicating a relative increase by a factor 1.8 in puff 2-48 C and a factor 2.2 in puff 4-81 B was attained in both puffs at 30 min after onset of puff induction.—Among staining procedures performed on sulphydryl groups, free α-amino acids and indole groups of tryptophane, only a positive result with the staining reaction on the indole groups was obtained for induced puffs.—Injection of tritiated sodium acetate, methionine-H3-methyl, ethionine-H3-ethyl, C14-sodium bicarbonate, a mixture of 15 H3-labelled L-amino acids and H3-tryptophane at various time intervals prior to puff induction failed to result in a specific incorporation of any of these radioactive substances into newly induced puffs.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alfert, M., Geschwind, J. J.: A selective staining method for the basic proteins of cell nuclei. Proc. nat. Acad. Sci. (Wash.) 39, 991–999 (1953).
Arnold, M.: Histochemie. Berlin-Heidelberg-New York: Springer 1968.
Beermann, W.: Chromomerenkonstanz und spezifische Modifikationen der Chromosomenstruktur in der Entwicklung und Organdifferenzierung von Chironomus tentans. Chromosoma (Berl.) 5, 139–198 (1952).
Benjamin, W. B., Goodman, R. M.: Phosphorylation of dipteran chromosomes and rat liver nuclei. Science 166, 629–630 (1969).
Berendes, H. D.: Salivary gland function and chromosomal puffing patterns in Drosophila hydei. Chromosoma (Berl.) 17, 35–77 (1965).
—: The hormone ecdysone as effector of specific changes in the pattern of gene activities of Drosophila hydei. Chromosoma (Berl.) 22, 274–293 (1967).
—: Factors involved in the expression of gene activity in polytene chromosomes. Chromosoma (Berl.) 24, 418–437 (1968).
—: Amino acid incorporation into giant chromosomes of Drosophila hydei. Dros. Inf. Serv. 42, 102 (1970).
—, Breugel, F. M. A. van, Holt, Th. K. H.: Experimental puffs in Drosophila hydei salivary gland chromosomes. Chromosoma (Berl.) 16, 35–47 (1965).
Breummer, N. C., Carver, M. J., Thomas, L. E.: A tryptophane histochemical method. J. Histochem. Cytochem. 5, 140–144 (1957).
—, Thomas, L. E.: Tryptophane histochemical method. J. Histochem. Cytochem. 6, 75 (1958).
Clever, U.: Genaktivitäten in den Riesenchromosomen von Chironomus tentans und ihre Beziehungen zur Entwicklung. I. Genaktivierungen durch Ecdyson. Chromosoma (Berl.) 12, 607–675 (1961).
—: Control of chromosome puffing. In: The control of nuclear activity (L. Goldstein, ed.), p. 161–186. New York: Prentice-Hall Inc. 1967.
Deitch, A. D.: Cytophotometry of proteins. In: Introduction to quantitative cytochemistry (G. L. Wied, ed.), p. 451–468. New York-London: Academic Press 1966.
—: Microspectrophotometric study of the binding of the anionic dye, naphthol yellow S by tissue sections and by purified proteins. Lab. Invest. 4, 324–351 (1955).
Ellgaard, E. G.: Gene activation without histone acetylation in Drosophila melanogaster. Science 157, 1070–1072 (1967).
Frenster, J. H.: Nuclear polyanions as de-repressors of synthesis of ribonucleic acid. Nature (Lond.) 206, 680–683 (1965).
Friedman, M., Shull, K. H., Farber, E.: Highly selective in vivo ethylation of rat liver nuclear protein by ethionine. Biochem. biophys. Res. Commun. 34, 857–864 (1969).
Gershey, E. L., Haslett, G. W., Vidali, G., Allfrey, V. G.: Chemical studies on histone methylation. Evidence for the occurence of 3-methyl histidine in avian erythrocyte histone fractions. J. biol. Chem. 244, 4871–4877 (1969).
Gorovsky, M. A., Woodard, J.: Histone-DNA ratios in five regions of a polytene chromosome of Drosophila virilis. J. Cell Biol. 31, 41 A (1966).
—: Histone content of chromosomal loci active and inactive in RNA synthesis. J. Cell Biol. 33, 723–728 (1967).
Kawamura, N.: Cytochemical and quantitative study of protein-bound sulfhydryl and disulphide groups in eggs of Arbacia during the first cleavage. Exp. Cell Res. 20, 127–128 (1960).
Keyl, H. G.: Duplikationen von Untereinheiten der chromosomalen DNS während der Evolution von Chironomus thummi. Chromosoma (Berl.) 17, 139–180 (1965).
Mazia, D., Brewer, P. A., Alfert, M.: The cytochemical staining and measurement of protein with mercuric bromphenol blue. Biol. Bull. (Woods Hole) 104, 57–67 (1953).
Pelling, C.: Ribonukleinsäure-Synthese der Riesenchromosomen. Autoradiographische Untersuchungen an Chironomus tentans. Chromosoma (Berl.) 15, 71–122 (1964).
Pettit, B. J., Rasch, R. W.: Tritiated-histidine incorporation into Drosophila salivary chromosomes. J. cell. comp. Physiol. 68, 325–334 (1966).
Pogo, B. G. T., Allfrey, V. G., Mirsky, A. E.: RNA synthesis and histone acetylation during the course of gene activation in lymphocytes. Proc. nat. Acad. Sci. (Wash.) 55, 805–812 (1966).
Rudkin, G. T.: Nucleic acid metabolism in giant chromosomes of Drosophila melanogaster. Ann. Histochim., Suppl. 2, 77–34 (1962).
Spelsberg, Th. C., Hnilica, L. S.: The effects of acidic proteins and RNA on the histone inhibition of the DNA-dependent RNA synthesis in vitro. Biochim. biophys. Acta (Amst.) 195, 63–75 (1969).
Swift, H.: Nucleic acids and cell morphology in dipteran salivary glands. In: The molecular control of cellular activity (J. Allen, ed.), p. 73–125. New York: McGraw-Hill 1962.
—: The histones of polytene chromosomes. In: The nucleohistones (J. Bonner and P. Ts'o, eds.), p. 169–183. San Francisco-London-Amsterdam: Holden Day Inc. 1964.
Teng, C., Hamilton, T. H.: Role of chromatin in estrogen action in the uterus. II. Hormone induced synthesis of non-histone acidic proteins which restore histone inhibited DNA-dependent RNA synthesis. Proc. nat. Acad. Sci. (Wash.) 63, 465–472 (1969).
Wang, T. Y.: Restoration of histone-inhibited DNA-dependent RNA synthesis by acidic chromatin proteins. Exp. Cell Res. 53, 288–291 (1968).
Wied, G. L.: Introduction to quantitative cytochemistry. New York-London: Academic Press 1966.
Yasuma, A., Itchikawa, T.: Ninhydrin-Schiff and alloxane-Schiff staining. A new histochemical staining method for protein. J. Lab. chim. Med. 41, 296–299 (1953).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Holt, T.K.H. Local protein accumulation during gene activation. Chromosoma 32, 64–78 (1970). https://doi.org/10.1007/BF00334011
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00334011