Changes in messenger RNA levels for the subunits of ribonucleotide reductase during the cell cycle of leukemia L1210 cells.
Ribonucleotide reductase consists of two non-identical protein subunits that are required for enzyme activity. These subunits are encoded by different genes and are not expressed coordinately as the cells pass through the cell cycle. Using specific cDNAs for the non-heme iron (NHI) and the effector-binding (EB) subunits the levels of the mRNAs for these two subunits were determined in leukemia L1210 cells during the transition from the G0/G1 phase to the S and G2/M phases of the cell cycle. Synchronized populations of L1210 cells were obtained either by ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) treatment or by enrichment by elutriation centrifugation. The changes in the levels of the mRNAs for NHI and EB subunits were compared with the changes in the levels of the mRNAs for actin, p53, c-myc, thymidine incorporation into DNA, and DNA content by flow cytometric measurements. Synchronization of the cells by the two methods resulted in quantitative differences in the responses. The EGTA synchronized L1210 cells showed maximal increases of 9.3- and 5.7-fold in the mRNAs for the NHI and EB subunits, respectively. The peak level of the NHI mRNA was observed at 12 hr after the addition of calcium ions. The peak increase in the level of the mRNA for the EB subunit was observed between 12 and 15 hr after the addition of calcium ions. The rate of increase for the mRNA for c-myc was greater than the increase in the mRNA for the NHI subunit.(ABSTRACT TRUNCATED AT 250 WORDS)
Thompson
D P
DP
Department of Internal Medicine, University of South Florida College of Medicine, H. Lee Moffitt Cancer Center and Research Institute, Tampa 33612.
Carter
G L
GL
Cory
J G
JG
D000818:Animals; D015152:Blotting, Northern; D002118:Calcium / Q000494:pharmacology; D002453:Cell Cycle; D004261:DNA Replication; D007700:Kinetics; D007939:Leukemia L1210 / Q000201:enzymology; D046911:Macromolecular Substances; D051379:Mice; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012264:Ribonucleotide Reductases / Q000235:genetics*; D013936:Thymidine / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology
D046911:Macromolecular Substances; D012333:RNA, Messenger; D012264:Ribonucleotide Reductases; D002118:Calcium; D013936:Thymidine