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-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).
Topp
H
H
Forschungsinstitut für Kinderernährung, Federal Republic of Germany.
Dörrenhaus
A
A
Bolt
H
H
Schöch
G
G
Föllmann
W
W
D000241:Adenosine / Q000031:analogs & derivatives* / Q000032:analysis; D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D011560:Pseudouridine / Q000032:analysis*; D012313:RNA / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D017382:Reactive Oxygen Species / Q000378:metabolism; D014529:Uridine / Q000031:analogs & derivatives* / Q000032:analysis
D012335:RNA, Ribosomal; D017382:Reactive Oxygen Species; D011560:Pseudouridine; C003268:N(6)-(N-threonylcarbonyl)adenosine; C010922:5,6-dihydrouridine; D012313:RNA; D012343:RNA, Transfer; D000241:Adenosine; D014529:Uridine