Abstract
Compartmentation of uridine 5′-triphosphate (UTP) was studied during the nucleolar synthesis of cytoplasmic ribosomal RNA (cyt-rRNA) and the synthesis of cytoplasmic transfer RNA (cyt-tRNA) in the nuclear matrix as well as the synthesis of mitochondrial ribosomal RNA (mt-rRNA) in tomato (Lycopersicon esculentum Mill. cv. Lukullus) cell-suspension culture using the approach of Wiegers et al. (Eur. J. Biochem. 64, 535–540, 1976). Before measurements were made, it was ensured that: (i) there was steady-state labeling of all RNAs studied as well as UTP; (ii) there was stability of cyt-tRNA and cyt-rRNA; (iii) there was no label randomization through degradation of [3H]uridine; (iv) there were significant differences in the specific radioactivity of UTP, the final immediate precursor of RNA, after supplying the cells with two different exogenous [3H]uridine concentrations.
By comparing the steady-state specific radioactivity of UTP with that of cyt-tRNA and cyt-18S rRNA during constant [3H]uridine supply, we found that the three molecules had equal specific radioactivities which, however, differed significantly from that of the mt-rRNA. With a 20-fold higher uridine concentration, i.e. a 20-fold lower specific radioactivity of exogenous [3H]uridine, the specific radioactivity of cyt-rRNA, cyt-tRNA and UTP decreased proportionally whereas that of mt-RNA increased. These results argue against different UTP pools during synthesis of cyt-rRNA and cyt-tRNA, but indicate compartmentation of UTP during rRNA synthesis in the nucleus and the mitochondria of tomato cells.
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Abbreviations
- CMP:
-
cytidine 5′-monophosphate
- cyt-rRNA:
-
cytoplasmic ribosomal RNA
- cyt-tRNA:
-
cytoplasmic transfer RNA
- mt-rRNA:
-
mitochondrial rRNA
- NC:
-
nitrocellulose
- PAGE:
-
polyacrylamide gel electrophoresis
- TLC:
-
thin-layer chromatography
- Tris:
-
2-amino-2-(hydroxymethyl)-1,3-propanediol
- UDP:
-
uridine 5′-diphosphate
- UMP:
-
uridine 5′-monophosphate
- UTP:
-
uridine 5′-triphosphate
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Hause, B., Wasternack, C. Compartmentation of uridine 5′-triphosphate occurs during synthesis of cytoplasmic and mitochondrial ribosomal RNAs of cultured tomato cells but not during synthesis of cytoplasmic ribosomal and transfer RNAs. Planta 176, 51–59 (1988). https://doi.org/10.1007/BF00392479
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DOI: https://doi.org/10.1007/BF00392479