Determination of degradation rates of transfer and ribosomal ribonucleic acids in cultured rat hepatocytes by measuring N6-threoninocarbonyladenosine, dihydrouridine, and pseudouridine in medium using high-performance liquid chromatography

Modified ribonucleic acid catabolites excreted into the medium by primary cultures of rat hepatocytes (2.3 +/- 0.42 x 10(6) cells/dish) during a 24-h cultivation period were quantified by reversed-phase high-performance liquid chromatography (fmol/10(3) cells): 613 +/- 81 dihydrouridine, 46 +/- 6 N6...

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Bibliographic Details
Published in:Analytical biochemistry 1997-12, Vol.254 (2), p.200-207
Main Authors: Topp, H, Dörrenhaus, A, Bolt, H, Schöch, G, Föllmann, W
Format: Article
Language:English
Subjects:
Adenosine - analogs & derivatives
Adenosine - analysis
Animals
Cells, Cultured
Chromatography, High Pressure Liquid
Liver - cytology
Liver - metabolism
Male
Pseudouridine - analysis
Rats
Rats, Inbred F344
Reactive Oxygen Species - metabolism
RNA - metabolism
RNA, Ribosomal - metabolism
RNA, Transfer - metabolism
Uridine - analogs & derivatives
Uridine - analysis
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Summary:Modified ribonucleic acid catabolites excreted into the medium by primary cultures of rat hepatocytes (2.3 +/- 0.42 x 10(6) cells/dish) during a 24-h cultivation period were quantified by reversed-phase high-performance liquid chromatography (fmol/10(3) cells): 613 +/- 81 dihydrouridine, 46 +/- 6 N6-threoninocarbonyladenosine, 1879 +/- 220 pseudouridine. On the basis of these excretion rates and the average frequency of occurrence of these modified ribonucleosides per cytoplasmic transfer ribonucleic acid (residues: 2.6 dihydrouridine, 0.22 N6-threoninocarbonyladenosine, 3 pseudouridine) as well as per cytoplasmic ribosomal ribonucleic acid (residues: 95 pseudouridine), the degradation rates of transfer and ribosomal ribonucleic acids were calculated. The degradation rate of transfer ribonucleic acid (fmol/10(3) cells/24 h) was 236 +/- 31 (via dihydrouridine) and 211 +/- 28 (via N6-threoninocarbonyladenosine) and that of ribosomal ribonucleic acid (fmol/10(3) cells/24 h) was 13.1 +/- 1.7 (via pseudouridine and N6-threoninocarbonyladenosine).
ISSN:0003-2697