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KMID : 0376219690060010033
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Chonnam Medical Journal
1969 Volume.6 No. 1 p.33 ~ p.44
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Ribonucleic Acid Synthesis in Nuclei from Chick Erythrocytes and Inhibitory Effect of NAD
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Abstract
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Evidence is presented indicating that the isolated nuclei from mature erythrocytes of chick still possess the capacity to synthesize RNA and that NAD exerts an inhibitory effect on this process.
1. Sucrose-washed nuclear preparations from saponin-hemolyzed mature erythrocytes were incubated with 32P-orthophosphate in the presence of glucose and KCI. After incubation RNA" was hydrolyzed with KOH, and the nucleotides in the alkaline hydrolysate were separated by anion-exchange chromatography. It was found that most of 32P incorporated into nuclear RNA was recovered in 2F´(3´)-nucleoside monophosphate indicating that 52P was actually incorporated into thephosphodiester bond of RNA. Tn contrast, a simple adsorption of 32P to the nuclear RNA and DNA was comparatively small.
2. This nuclear RNA synthesizing system showed a DNA -dependence but did not regiure Mg2{´ and preformed nucleoside triphosphates.
3. Incorporation of 32P into the alkaline digest of RNA was depressed byNAD. The inhibition by NAD was roughly proportional to the concentr ation of NAD {0.05-1.0 mM), and released when-nicotinamide or heroin was added togetherthe situation being very similar to the inhibitory effect o NAD on-the nuclear protein synthesis previously observed. Poly ADP-ribose --also inhibited markedly the nuclear ,RNA synthesis. However, neither DNA from chick erythrocyte nuclei nor ribosomal RNA of rabbit reticulocytes exerted any significant effect.
4. Chick erythrocyte nuclei also incorporated 32PO4 into nucleotides suggesting that before being incorporated into RNA
incorporated into nucleoside triphosphates as is the case for RNA occurring in other animal cell nuclei.
5. Nuclear ribonuclease and adenosinetriphosphatase were also demonstrable in chick erythrocyte nuclei.
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