Abstract
Abnormal cell cycle regulation is believed to be an important step in tumorigenesis. In mammalian cells, DNA damage commonly leads to cell cycle arrest in G2; however, little is known about the detailed biochemical mechanisms underlying the DNA damage‐induced G2 arrest. In order to identify genes differentially expressed in association with G2 arrest, differential display analysis was performed between exponentially growing Chinese hamster ovary (CHO) cells and G2‐arrested CHO cells induced by etoposide, SN‐38, or X‐radiation. We identified five cDNA clones whose expression was up‐regulated in G2‐arrested CHO cells. Sequence analysis revealed that three clones were homologous to known genes: isogene I of translation initiation factor eIF‐4A, ribosomal protein L13, and translation repressor NAT1. The remaining two clones showed no homology to known genes. These results indicate that DNA damage can alter the expression of multiple genes, including translational regulators. J. Clin. Lab. Anal. 14: 314–319, 2000. © 2000 Wiley‐Liss, Inc.
Keywords: cell cycle, G2/M, differential display, eIF‐4A, ribosomal protein L13, NAT1
REFERENCES
- 1. Smith ML, Fornace AJ Jr. 1996. Mammalian DNA damage‐inducible genes associated with growth arrest and apoptosis. Mutat Res 340:109–124. [DOI] [PubMed] [Google Scholar]
- 2. Kastan MB, Onyekwere O, Sidransky D, Vogelstein B, Craig RW. 1991. Participation of p53 protein in the cellular response to DNA damage. Cancer Res 51:6304–6311. [PubMed] [Google Scholar]
- 3. el‐Deiry WS, Tokino T, Velculescu VE, et al. 1993. WAF1, a potential mediator of p53 tumor suppression. Cell 75:817–825. [DOI] [PubMed] [Google Scholar]
- 4. Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R, Beach D. 1993. p21 is a universal inhibitor of cyclin kinases. Nature 366:701–704. [DOI] [PubMed] [Google Scholar]
- 5. Harper JW, Adami GR, Wei N, Keyomarsi K, Elledge SJ. 1993. The p21 Cdk‐interacting protein Cip1 is a potent inhibitor of G1 cyclin‐ dependent kinases. Cell 75:805–816. [DOI] [PubMed] [Google Scholar]
- 6. Draetta G, Beach D. 1988. Activation of cdc2 protein kinase during mitosis in human cells: cell cycle‐dependent phosphorylation and subunit rearrangement. Cell 54:17–26. [DOI] [PubMed] [Google Scholar]
- 7. Sanchez Y, Wong C, Thoma RS, et al. 1997. Conservation of the Chk1 checkpoint pathway in mammals: linkage of DNA damage to Cdk regulation through Cdc25. Science 277:1497–1501. [DOI] [PubMed] [Google Scholar]
- 8. Peng CY, Graves PR, Thoma RS, Wu Z, Shaw AS, Piwnica‐Worms H. 1997. Mitotic and G2 checkpoint control: regulation of 14‐3‐3 protein binding by phosphorylation of Cdc25C on serine‐216. Science 277:1501–1505. [DOI] [PubMed] [Google Scholar]
- 9. Hermeking H, Lengauer C, Polyak K, et al. 1997. 14‐3‐3 sigma is a p53‐regulated inhibitor of G2/M progression. Mol Cell 1:3–11. [DOI] [PubMed] [Google Scholar]
- 10. Hartwell L. 1992. Defects in a cell cycle checkpoint may be responsible for the genomic instability of cancer cells. Cell 71:543–546. [DOI] [PubMed] [Google Scholar]
- 11. Paulovich AG, Toczyski DP, Hartwell LH. 1997. When checkpoints fail. Cell 88:315–321. [DOI] [PubMed] [Google Scholar]
- 12. Hartwell LH, Kastan MB. 1994. Cell cycle control and cancer. Science 266:1821–1828. [DOI] [PubMed] [Google Scholar]
- 13. Liang P, Pardee AB. 1992. Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257:967–971. [DOI] [PubMed] [Google Scholar]
- 14. Chomczynski P, Sacchi N. 1987. Single‐step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Anal Biochem 162:156–159. [DOI] [PubMed] [Google Scholar]
- 15. Helps NR, Adams SM, Brammar WJ, Varley JM. 1995. The Drosophila melanogaster homologue of the human BBC1 gene is highly expressed during embryogenesis. Gene 162:245–248. [DOI] [PubMed] [Google Scholar]
- 16. Olvera J, Wool IG. 1994. The primary structure of rat ribosomal protein L13. Biochem Biophys Res Commun 201:102–107. [DOI] [PubMed] [Google Scholar]
- 17. Saez‐Vasquez J, Raynal M, Meza‐Basso L, Delseny M. 1993. Two related, low‐temperature‐induced genes from Brassica napus are homologous to the human tumour bbc1 (breast basic conserved) gene. Plant Mol Biol 23:1211–1221. [DOI] [PubMed] [Google Scholar]
- 18. Adams SM, Helps NR, Sharp MG, Brammar WJ, Walker RA, Varley JM. 1992. Isolation and characterization of a novel gene with differential expression in benign and malignant human breast tumours. Hum Mol Genet 1:91–96. [DOI] [PubMed] [Google Scholar]
- 19. Grifo JA, Tahara SM, Leis JP, Morgan MA, Shatkin AJ, Merrick WC. 1982. Characterization of eukaryotic initiation factor 4A, a protein involved in ATP‐dependent binding of globin mRNA. J Biol Chem 257:5246–5252. [PubMed] [Google Scholar]
- 20. Ford MJ, Anton IA, Lane DP. 1988. Nuclear protein with sequence homology to translation initiation factor eIF‐4A. Nature 332:736–738. [DOI] [PubMed] [Google Scholar]
- 21. Nielsen PJ, Trachsel H. 1988. The mouse protein synthesis initiation factor 4A gene family includes two related functional genes which are differentially expressed. EMBO J 7:2097–2105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22. Williams‐Hill DM, Duncan RF, Nielsen PJ, Tahara SM. 1997. Differential expression of the murine eukaryotic translation initiation factor isogenes eIF4AI and eIF4AII is dependent upon cellular growth status. Arch Biochem Biophys 338:111–120. [DOI] [PubMed] [Google Scholar]
- 23. Yamanaka S, Poksay KS, Arnold KS, Innerarity TL. 1997. A novel translational repressor mRNA is edited extensively in livers containing tumors caused by the transgene expression of the apoB mRNA‐editing enzyme. Genes Dev 11:321–333. [DOI] [PubMed] [Google Scholar]
- 24. Smith PJ, Anderson CO, Watson JV. 1986. Predominant role for DNA damage in etoposide‐induced cytotoxicity and cell cycle perturbation in human SV40‐transformed fibroblasts. Cancer Res 46:5641–5645. [PubMed] [Google Scholar]
- 25. Kawato Y, Aonuma M, Hirota Y, Kuga H, Sato K. 1991. Intracellular roles of SN‐38, a metabolite of the camptothecin derivative CPT‐11, in the antitumor effect of CPT‐11. Cancer Res 51:4187–4191. [PubMed] [Google Scholar]
- 26. Radford IR. 1986. Effect of radiomodifying agents on the ratios of X‐ray‐induced lesions in cellular DNA: use in lethal lesion determination. Int J Radiat Biol Relat Stud Phys Chem Med 49:621–637. [DOI] [PubMed] [Google Scholar]
- 27. Brenner DJ, Ward JF. 1992. Constraints on energy deposition and target size of multiply damaged sites associated with DNA double‐strand breaks. Int J Radiat Biol 61:737–748. [DOI] [PubMed] [Google Scholar]
