Summary
DNA binding proteins (DBP) are shown to be engaged in the regulatory functions of the genome. Since the accelerated epidermopoesis in psoriasis indicates changed gene activities, the examination of DBP possibly could help to clarify the pathological processes. Psoriatic scales were used to elaborate the method. DBP was isolated from a protein extract by affinity chromatography with DNA-cellulose according to Alberts et al. About 3-5% of postribosomal crude protein (PCP) extract had an affinity to DNA. The separation of DBP on SDS polyacrylamide slab gels showed a characteristic spectrum of proteins with molecular weights from 12.7 to 150 × 103 daltons. The protein band spectrum of the concentrated DBP fraction was different from that of the crude extract. These observations show the isolated DBP to represent an own group of water soluble proteins with DNA binding abilities.
Zusammenfassung
Bei der Aufklärung von Regulationsvorgängen am Genom wurden die chromosomalen DNA-affinen Proteine (DBP) als wesentliche Faktoren erkannt. Da die gesteigerte Epidermopoese bei der Psoriasis auf veränderte Genaktivitäten hinweist, könnten Untersuchungen der DBP ein Weg zur molekularbiologischen Klärung der pathologischen Vorgänge bei dieser Dermatose sein. Zur Ausarbeitung der Methodik dienten Psoriasisschuppen. Die Isolierung der DBP erfolgte aus einem Proteinextrakt affinitätschromatographisch mit DNA-Cellulose nach Alberts et al. Unter den Versuchsbedingungen besaßen 3-5% der Rohextraktproteine Affinität zur DNA. Die Auftrennung der DBP im SDS-Plattengel ergab ein charakteristisches Spektrum an Proteinen mit Molekulargewichten von 12700–150000. Das Bandenspektrum der angereicherten DBP-Fraktion unterschied sich hierbei von dem des Rohextraktes. Somit stellen die DBP eine eigene Gruppe an löslichen Proteinen mit erhaltener DNA-Affinität dar.
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References
Alberts, B. M., Amadio, F. J., Jenkins, M., Gutmann, E. D., Ferris, F. L.: Studies with DNA-cellulose chromatography. I. DNA-binding proteins from E. coli. Cold Spring Harbour Symp. Quant. Biol.33, 289–304 (1968)
Burton, K.: Biochem. J.62, 315 (1956)
Chae, C.-B.: Reconstitution of chromatin-mode of reassociation of chromosomal proteins. Biochem.14, 900–906 (1975)
Dewair, M. A., Matthaei, H.: Tissue specificity of non-histone proteins from human chromatin. Molec. gen. Genet.146, 309–312 (1976)
Dunker, A. K., Rueckert, R. R.: Observations of molecular weight determinations on polyarylamid gel. J. Biol. Chem.244, 5074–5080 (1969)
Fellenberg, G.: Chromosomale proteine, p. 27–57. Stuttgart: E. Ulmer 1974
Francksen, H., Garadi, R.: Complete recovery of proteins in concentrators and some drawlochs, revealed by polyacrylamide gel electrophoresis. Z. Analyt. Chem.271, 340–344 (1974)
Jost, E., Lennox, R., Harris, H.: Affinity chromatography of DNA-binding proteins from human, murine and man-mouse hybrid cell lines. J. Cell Sci.18, 41–65 (1975)
Kostraba, N. C., Montagna, R. M., Wang, T. Y.: Study of the loosely bound non-histone chromatin proteins. J. Biol. Chem.250, 1548–1555 (1975)
Kostraba, N. C., Wang, T. Y.: Inhibition of transcription in vitro by a non-histone protein isolated from Ehrlich ascites tumor chromatin. J. Biol. Chem.250, 8938–8942 (1975)
Laemmli, U. K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T 4. Nature (Lond.)227, 680–685 (1970)
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Kolorimetrische Proteinbestimmung mit dem Folinschen Phenolreagenz. J. Biol. Chem.193, 265 (1951)
Mainwaring, W. I. P., Renni, P. S., Keekn, Y.: The androgenic regulation of prostate proteins with a high affinity for DNA. Biochem. J.156, 253–264 (1976)
Peter, G., Brand, A., Peter, R.: Studies on the species, amount and base composition of nucleic acids in psoriatic scales and normal horny layer. Dermatologica (Basel)151, 333–341 (1975)
Reimer, G., Ramb, G., Leonhardi, G.: Isolation and physico-chemical characterisation of the DNA from normal human skin and psoriatic scales. Arch. Derm. Res.256, 241–246 (1976)
Stein, G. H.: Differences in DNA-binding proteins isolated from normal and transformed human cells. Exp. Cell Res.99, 115–125 (1976)
Stein, G., Kleinsmith, L. J.: Chromosomal proteins and their role in the regulation of gene expression. New York: Academic Press 1976
Tashiro, L., Kurokowa, S.: Chromatin conformation as affected by shearing and by the removal of a portion of non-histone proteins. Febs Letters59, 250–253 (1975)
Teng, C. S., Teng, C. T., Allfrey, U. G.: Studies of nuclear proteins. J. Biol. Chem.246, 3597–3609 (1971)
Weber, K., Osborn, M.: The reliability of molecular weight determination by SDS-polyacrylamide gel electrophoresis. J. Biol. Chem.244, 4406–4412 (1969)
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This work was supported by a grant of the Deutsche Forschungsgemeinschaft
We are greatly indebted to Mrs. M. Mentzer for her competent assistance
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Reimer, G., Leonhardi, G. & Schwulera, V. DNA-Binding proteins of psoriatic scales. Arch. Derm. Res. 260, 39–46 (1977). https://doi.org/10.1007/BF00558012
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DOI: https://doi.org/10.1007/BF00558012