Summary
DNA-bindung proteins (DBP) of normal human dermis, epidermis, horny layer and psoriatic scales represent a tissue-specific group of mostly nuclear nonhistone proteins. To analyse their function, the different DBP fractions were examined concerning the presence of DNase, DNA-polymerase and RNA-polymerase activities.
DBP of normal epidermis and horny layer contain four different DNases. One DNase of both DBP fractions is active only at pH 5.0. Three DNases of epidermal DBP are active at a pH-range from 5.0–8.5, while the corresponding DNases of horny layer-DBP are most active at pH 7.4. Probably these DNases have changed their pH-optimum during keratinisation. DBP of psoriatic scales include no activity of these three DNases and the pH 5.0-DNases seem to have reduced DNA-affinity. Human dermis DBP contain quite another set of four DNases which hardly can be correlated to the DNases of epidermal DBP.
DNA-polymerase activities are present in each fraction and derive from different DNA-polymerases. Two DNA-polymerases with pI-values of 4.5 and 9.3 may correspond to β- and α-DNA-polymerase of eukaryotes, respectively. Further activity of proteins which are focussed at pH 6.5–7.2 and 8.2 could be detected. The proteins represent either tissue-specific DNA-polymerases or further thymidine monophosphate incorporating enzymes. Contrary, RNA-polymerase activity could not be enriched from correlating extracts by DNA-cellulose chromatography.
Zusammenfassung
DNA-affine Proteine (DBP) aus normaler menschlicher Haut, Epidermis, Hornschicht und Psoriasisschuppen sind eine gewebsspezifische Proteingruppe, die zumeist aus nukleären Nichthiston-Proteinen besteht. Um ihre Funktion zu analysieren, wurden sie auf enzymatische Aktivitäten von DNase, DNA-Polymerase und RNA-Polymerase untersucht.
DBP der Epidermis und der Hornschicht enthalten vergleichbare Aktivitäten von vier verschiedenen DNasen. Eine davon ist nur bei pH 5.0 aktiv. Die anderen DNasen scheinen während der Keratinisierung ihr pH-Optimum zu verändern: DNasen der Epidermis-DBP sind aktiv im pH-Bereich von 5.0–8.5, während die entsprechenden DNasen der Hornschicht-DBP bei pH 7.4 ihre größte Aktivität zeigen. In Psoriasisschuppen sind diese DNasen inaktiv, während die pH 5.0-DNasen offenbar eine verringerte DNA-Affinität besitzen. DBP der menschlichen Haut enthalten Aktivitäten von vier anderen DNasen, die nicht mit den DNasen epidermaler DBP vergleichbar sind.
DNA-Polymerase-Aktivität ist in jeder DBP-Fraktion enthalten und beruht auf verschiedenen DNA-Polymerasen. Zwei DNA-Polymerasen mit pI-Werten von 4.5 und 9.3 dürften der eukaryotischen β-bzw. α-DNA-Polymerase entsprechen. Weiterhin wurden DNA synthetisierende Proteine mit pI-Werten von 6,5–7,2 und 8,2 gefunden, die entweder gewebsspezifische DNA-Polymerasen oder andere Thymidinmonophosphat einbauende Enzyme darstellen. RNA-Polymerasen konnten aus den entsprechenden Rohextrakten nicht durch DNA-Affinitätschromatographie angereichert werden.
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Abbreviations
- (d)ATP:
-
(deoxy)adenosine 5′-triphosphate
- (d)CTP:
-
(deoxy)cytine 5′-triphosphate
- DBP:
-
DNA-binding proteins
- DNase:
-
deoxyribonucleate 5′-oligonucleotidohydrolase (EC 3.1.4.5) = deoxyribonuclease
- EDTA:
-
ethylendiaminetetraacetic acid sodium salt
- (d)GTP:
-
(deoxy)guanosine 5′-triphosphate
- RNase:
-
ribonucleate 5′-oligonucleotidohydrolase (EC 3.1.4.22) = ribonuclease
- TCA:
-
trichloroacetic acid
- TTP:
-
thymidine 5′-triphosphate
- TMP:
-
thymidine 5′-monophosphate
- UTP:
-
uridine 5′ triphosphate
- UMP:
-
uridine 5′ monophosphate
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Reimer, G., Zöllner, E.J., Reitz, M. et al. Comparison of DNase, DNA-polymerase and RNA-polymerase activities present in the DNA-bindung proteins of normal human dermis, epidermis, horny layer and psoriatic scales. Arch Dermatol Res 263, 317–324 (1978). https://doi.org/10.1007/BF00446948
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DOI: https://doi.org/10.1007/BF00446948