RNA Synthesis and polysome formation in pollen tubes
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The formation of polysomes in relation to RNA synthesis was investigated in tobacco (Nicotiana tabacum L.) pollen cultivated submersely for a period of 12 h. The percentage of polysomes was estimated by determining the number of ribosomes carrying nascent polypeptides using RNase and 0.8 M KC1 treatment of the ribosomal preparation. This approach showed that the proportion of ribosomes “active” in protein synthesis amounts to about 12 % in dry pollen rising to 46 % within 10 min of imbibition and to 66 % during the period 1–4 h of cultivation. The latter increase is accompanied by a rapid incorporation of uridine-14C into polysome-as-sociated RNA and is sensitive to actinomycin D.
The rapidly labelled RNA isolated from the ribosomal preparation sedirnented in the range from about 5S to 30S. Longer labelling periods led to a gradual shift of the major peak of this heterogeneous RNA from about 11.5S and 14S to about 7.5S and the development of radioactivity peaks in the position of 18S and 28S RNA. Uridine incorporated both into the active ribosomes and into the subunits of inactive ribosomes at a more or less constant rate during the whole 12-h period of cultivation.
These results present evidence that in addition to the initial combination of the existing ribosomes with the stored mRNA following imbibition, an activation of pollen tube genome and its implication in directing protein synthesis take place during the early phases of pollen tube growth. From the results on kinetics of labelling of ribosomes it appears that in pollen tubes new synthesized ribosomal subunits enter polysomes directly, the entry of 40S subunits being more rapid than that of 60S subunits.
KeywordsRibosomal Subunit Nascent Polypeptide Sucrose Density Gradient Centrifugation Jerusalem Artichoke Tuber Active Ribosome
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