Changes in the rate of RNA synthesis during the cell cycle.

Abstract

Although the rate of RNA synthesis is known to drop at mitosis, the recent identification of 11 stages in the cell cycle (El-Alfy et al., 1994) makes it possible to measure the rate of this synthesis at each one of the stages and thus find out how it varies throughout the cell cycle. Mice were injected intravenously with the RNA precursor, 3H-uridine; the duodenum was fixed 5-15 minutes later for embedment in Epon, and the duodenal crypts were cut in semithin serial sections for study of the rapidly dividing crypt columnar cells. Using Feulgen-stained sections, each cell nucleus was assigned to one of the 11 stages described in the cell cycle, and the same nucleus was identified in the next serial section that had been processed for radioautography, so that the overlying silver grains were enumerated. The count was taken as an index of the rate of RNA synthesis by this nucleus. Starting from stage I of the cell cycle (the period defined by the presence of a minimal amount of chromatin during which the S phase begins) and up to stage IV (when the S phase ends and the G2 phase begins), all or nearly all nuclei are synthesizing RNA with the rate peaking at stage III. During stages V to VIII (the period comprising the mitotic steps), the percentage of RNA-synthesizing nuclei decreases to over half at stage V (prophase), -10% at stages VIa (prometaphase) and VIb (metaphase) and none at stages VII (anaphase) and VIII (telophase). During stages IX-XI (which correspond to the G1 phase), the percentage rises sharply at stage XI to reach up to 100% at stages X and XI. Finally, on the average, 35% of nuclear silver grains are over the nucleolus (presumably representing ribosomal RNA precursors), whereas 65% are over the nucleoplasm (presumably representing mainly heterogeneous RNA precursors). Cells synthesize RNA during the interphase, but at a variable rate with a peak in S. The synthesis proceeds in a majority of the cells at prophase, but only in a few of them at prometaphase and metaphase, and in none at anaphase and telophase.