Zusammenfassung
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1.
Der Abbau der RNS in toluolbehandelten stationären Zellen von Escherichia coli ist qualitativ und quantitativ sehr ähnlich dem Abbau in zellfreien Extrakten und isolierten Ribosomen. Er ist abhängig von pH-Wert und Temperatur.
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2.
In toluolbehandelten Zellen gibt es zwei Wege des RNS-Zerfalls, die für zellfreie Extrakte und Ribosomen bereits bekannt sind: In TrisÄDTA-Puffer entstehen 2′-3′-cyclische Phosphate, in Tris-Mg-Na2HPO4-Puffer 5′-Nucleotide und 5′-Nucleosiddiphosphate. Die Nucleotide werden weiter in Nucleoside und/oder Basen zerlegt.
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3.
Durch Versuche mit RNase I-losen Stämmen konnte gezeigt werden, daß im Tris-ÄDTA-Medium nur die RNase I wirksam ist. Der Abbau zu 5′-Phosphaten wird vermutlich wie in zellfreien Extrakten oder Ribosomen über eine Polynucleotidphosphorylase hervorgerufen.
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4.
Auch nach der Zerstörung der cytoplasmatischen Membran von E. coli durch Phenol, Phenyläthylalkohol, Isopropanol, Isoamylalkohol, Allylisothiocyanat, Polymyxin B oder Gefriertrocknung wird ein RNase I-bedingter RNS-Abbau beobachtet.
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5.
In toluolbehandelten Zellen von Alcaligenes faecalis läßt sich keine aktive RNase I nachweisen.
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6.
Der Mechanismus, der zur RNase I-Wirkung führt, wird diskutiert.
Summary
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1.
The degradation of RNA in toluenized stationary cells of Escherichia coli is quantitatively and qualitatively very similar to the degradation in cell-free extracts and isolated ribosomes. This depolymerization depends on pH and temperature.
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2.
In toluenized cells there are two possible routes of RNA-degradation as known in cell-free extracts and ribosomes: In Tris-EDTA buffer 2′,3′-cyclic nucleotides and in Tris-Mg-Na2HPO4 medium 5′-phosphates and 5′-diphosphates are found as degradation products. The nucleotides are further split into nucleosides and/or bases.
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3.
Using RNase I-less strains it could be verified that in Tris-EDTA buffer only RNase I is active in degrading ribosomes of toluenized cells. The depolymerization of ribosomal RNA into 5′-phosphates may be caused as in cell-free extracts and ribosomes by a polynucleotide phosphorylase.
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4.
After membrane injury in E. coli by phenol, phenethyl alcohol, isopropyl alcohol, isoamyl alcohol, allylisothiocyanate, polymyxin B and freeze drying RNase I-induced RNA-degradation is observed too.
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5.
In toluene treated cells of Alcaligenes faecalis no RNase I-activity is detectable.
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6.
The mode of action of RNase I in membrane damaged bacteria is discussed.
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Teuber, M. Bedingungen für den Abbau von Ribonucleinsäure in Escherichia coli nach Zerstörung der cytoplasmatischen Membran durch Toluol. Archiv. Mikrobiol. 55, 31–45 (1966). https://doi.org/10.1007/BF00409154
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DOI: https://doi.org/10.1007/BF00409154