Cell-cycle-directed regulation of thymidylate synthase messenger RNA in human diploid fibroblasts stimulated to proliferate.
Human diploid fibroblasts were synchronized in the resting phase by incubation in medium containing a low level of serum and then stimulated to proliferate by adding a high concentration of serum. DNA replication started 12 hours after addition of serum, and reached a maximum after 24 hours. Thymidylate synthase activity was very low in resting cells, but began to increase 12 hours after growth stimulation and thereafter continued to increase. Thymidylate synthase mRNA in the growing cells was compared with that in resting cells, using cloned human thymidylate synthase cDNA as a probe. Results showed that the mRNA content as a percentage of total RNA began to increase six hours after stimulation, reaching a level about 14 times that in unstimulated cells after 24 hours. However, the mRNA content relative to poly(A)+ RNA had increased two- to fourfold by 24 hours after growth stimulation. Transcription of the thymidylate synthase gene, determined by hybridizing labelled nascent transcripts obtained in isolated nuclei to immobilized human thymidylate synthase cDNA, was similar in the nuclei of resting and of growth-stimulated cells. These results show that the increase in thymidylate synthase mRNA in growth-stimulated cells is caused by an increase in post-transcriptional events.
D002455:Cell Division; D004247:DNA / Q000096:biosynthesis; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D005786:Gene Expression Regulation; D006801:Humans; D009693:Nucleic Acid Hybridization; D011061:Poly A / Q000378:metabolism; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D013940:Thymidylate Synthase / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D004247:DNA; D013940:Thymidylate Synthase