Determination of queuosine derivatives by reverse-phase liquid chromatography for the hypomodification study of Q-bearing tRNAs from various mammal liver cells.
Three queuosine derivatives (Q-derivatives) have been found at position 34 of four mammalian so-called Q-tRNAs: queuosine (Q) in tRNA(Asn) and tRNA(His), mannosyl-queuosine (manQ) in tRNA(Asp), and galactosyl-queuosine (galQ) in tRNA(Tyr). An analytical procedure based on the combined means of purified tRNA isolation from liver cells and ribonucleoside analysis by reverse-phase high performance liquid chromatography coupled with real-time UV-spectrometry (RPLC-UV) was developed for the quantitative analysis of the three Q-derivatives present in total tRNA from liver tissues and liver cell cultures. Using this analytical procedure, the rates of Q-tRNA modification were studied in total tRNAs from various mammalian hepatic cells. Our results show that the four Q-tRNAs are fully modified in liver tissues from adult mammals, regardless of the mammal species. However, a lack in the Q-modification level was observed in Q-tRNAs from newborn rat liver, as well in Q-tRNAs from normal rat liver cell cultures growing in a low queuine content medium, and from a rat hepatoma cell line. It is noteworthy that in all cases of Q-tRNA hypomodification, our analytical procedure showed that tRNA(Asp) is always the least affected by the hypomodification. The biological significance of this phenomenon is discussed.
Costa
Annie
A
Faculté de Médecine et Centre Hospitalier Universitaire, Université de Bourgogne, 10 Bd de-Lattre-de-Tassigny, 21079 Cedex, Dijon, France.
Païs de Barros
Jean Paul
JP
Keith
Gérard
G
Baranowski
Wlodzimierz
W
Desgrès
Jean
J
D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D002851:Chromatography, High Pressure Liquid; D022781:Hepatocytes / Q000737:chemistry; D008099:Liver / Q000737:chemistry*; D008114:Liver Neoplasms, Experimental; D009704:Nucleoside Q / Q000031:analogs & derivatives* / Q000032:analysis*; D012343:RNA, Transfer / Q000737:chemistry* / Q000302:isolation & purification; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry; D012348:RNA, Transfer, Asn / Q000737:chemistry; D051381:Rats; D014407:Tumor Cells, Cultured
D012346:RNA, Transfer, Amino Acyl; D012348:RNA, Transfer, Asn; D009704:Nucleoside Q; D012343:RNA, Transfer