Characterization of single-stranded regions in DNA from injured tissue

• Published in: Experimental and molecular pathology Vol. 41; no. 1; pp. 1 - 9
• Main Authors: Terry Warnick, C., Dierenfeldt, Susan M., Lazarus, Harrison M.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.08.1984; Elsevier
• Subjects: Ageing, cell death; Animals; Base Composition; Biological and medical sciences; Cell physiology; Cold Temperature; DNA, Single-Stranded - metabolism; Endonucleases - metabolism; Fundamental and applied biological sciences. Psychology; Hot Temperature; Hydrolysis; In Vitro Techniques; Ischemia - metabolism; Kidney - blood supply; Male; Mice; Molecular and cellular biology; Single-Strand Specific DNA and RNA Endonucleases; Time Factors; Tissue; Temperature; Cold; Cell death; DNA; Shelf life; Molecular weight; Kidney
• ISSN: 0014-4800; 1096-0945
• DOI: 10.1016/0014-4800(84)90002-9

The objective of this study was to characterize the previously demonstrated decrease in the molecular weight of DNA from kidneys submitted to storage injury. DNA from kidneys stored at either 0°C for 24–96 hr or 37°C for 1–3 hr underwent limited hydrolysis when incubated with S 1 nuclease, an enzyme which specifically degrades single-stranded DNA. For warm-storage injury (37°C), the susceptibility of the DNA toward S 1 nuclease hydrolysis increased progressively with storage time. In the case of cold-storage injury (0°C), a maximum degree of single strandedness was observed in the DNA representing 60 hr of storage. DNA from kidneys stored for 72 hr or longer was a poor substrate for S 1 nuclease. Additionally, the nucleotide composition of the single-stranded regions was analyzed by high-performance liquid chromatography (HPLC). The results showed that single-stranded regions initially rich in dA and dT are formed during warm-storage injury. No such favoritism for a particular base was observed in single-stranded regions produced during cold-storage injury. The data suggest that both warm- and cold-storage injury promote DNA degradation. The storage temperature apparently dictates the mechanism(s) by which the degradative process proceeds.