Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Nov 11;20(21):5527–5531. doi: 10.1093/nar/20.21.5527

DNA damage response of cloned DNA beta-polymerase promoter is blocked in mutant cell lines deficient in protein kinase A.

E W Englander 1, S H Wilson 1
PMCID: PMC334382  PMID: 1454516

Abstract

DNA beta-polymerase (beta-pol), one of the recognized DNA polymerizing enzymes in vertebrates, has a role in 'very short patch' gap-filling synthesis during nucleotide excision DNA repair. In human and mouse, the enzyme is encoded by a single-copy gene located on the short arm of chromosome 8 near the centromere. In a series of studies, we have found that the cloned human beta-pol promoter is regulated by signals acting through the single ATF/CRE palindrome in the core promoter. These signals include transactivation by: adenovirus E1a/E1b proteins; activated p21ras; and in CHO cells, treatment with the DNA damaging agent MNNG. Hence, several types of stimulatory signals are mediated through the single ATF/CRE site, including DNA damage induction. To understand the mechanism of beta-pol promoter activation by MNNG in CHO cells, we asked whether induction of the cAMP/protein kinase A pathway can increase transcription of the cloned promoter in this system. Agents that increase cellular cAMP levels (8-BrcAMP; forskolin and IBMx) activated the beta-pol promoter fusion gene in transient expression experiments, and a mutation in the ATF/CRE palindrome blocked this response. Thus, the ATF/CRE site appears to be cAMP responsive in the CHO cell system. We found that the activation of the cloned beta-pol promoter by MNNG does not occur with two mutant CHO cell lines that are deficient in protein kinase A activity. Further, simultaneous treatment of wild-type CHO cells, with MNNG and to elevate cAMP, failed to result in an additive effect for activation of the beta-pol promoter. Thus, these effectors may act through a common pathway. These results suggest that the activation of the cloned beta-pol promoter in CHO cells following MNNG treatment is mediated through the cAMP/protein kinase A signal transduction pathway.

Full text

PDF
5527

Images in this article

5528Image
on p.5528
5528Image
on p.5528
5529Image
on p.5529
5529Image
on p.5529

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abraham I., Brill S., Hyde J., Fleischmann R., Chapman M., Gottesman M. M. DNA-mediated gene transfer of a mutant regulatory subunit of cAMP-dependent protein kinase. J Biol Chem. 1985 Nov 15;260(26):13934–13940. [PubMed] [Google Scholar]
  2. Büscher M., Rahmsdorf H. J., Litfin M., Karin M., Herrlich P. Activation of the c-fos gene by UV and phorbol ester: different signal transduction pathways converge to the same enhancer element. Oncogene. 1988 Sep;3(3):301–311. [PubMed] [Google Scholar]
  3. Chang L. M. Low molecular weight deoxyribonucleic acid polymerase from calf thymus chromatin. I. Preparation of homogeneous enzyme. J Biol Chem. 1973 Jun 10;248(11):3789–3795. [PubMed] [Google Scholar]
  4. Chang L. M. The distributive nature of enzymatic DNA synthesis. J Mol Biol. 1975 Apr 5;93(2):219–235. doi: 10.1016/0022-2836(75)90129-1. [DOI] [PubMed] [Google Scholar]
  5. Devary Y., Gottlieb R. A., Lau L. F., Karin M. Rapid and preferential activation of the c-jun gene during the mammalian UV response. Mol Cell Biol. 1991 May;11(5):2804–2811. doi: 10.1128/mcb.11.5.2804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dresler S. L., Lieberman M. W. Identification of DNA polymerases involved in DNA excision repair in diploid human fibroblasts. J Biol Chem. 1983 Aug 25;258(16):9990–9994. [PubMed] [Google Scholar]
  7. Englander E. W., Widen S. G., Wilson S. H. Mammalian beta-polymerase promoter: phosphorylation of ATF/CRE-binding protein and regulation of DNA binding. Nucleic Acids Res. 1991 Jun 25;19(12):3369–3375. doi: 10.1093/nar/19.12.3369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Englander E. W., Wilson S. H. Protein binding elements in the human beta-polymerase promoter. Nucleic Acids Res. 1990 Feb 25;18(4):919–928. doi: 10.1093/nar/18.4.919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Englander E. W., Wilson S. H. The cloned promoter of the human DNA beta-polymerase gene contains a cAMP response element functional in HeLa cells. DNA Cell Biol. 1992 Jan-Feb;11(1):61–69. doi: 10.1089/dna.1992.11.61. [DOI] [PubMed] [Google Scholar]
  10. Fornace A. J., Jr, Zmudzka B., Hollander M. C., Wilson S. H. Induction of beta-polymerase mRNA by DNA-damaging agents in Chinese hamster ovary cells. Mol Cell Biol. 1989 Feb;9(2):851–853. doi: 10.1128/mcb.9.2.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gonzalez G. A., Montminy M. R. Cyclic AMP stimulates somatostatin gene transcription by phosphorylation of CREB at serine 133. Cell. 1989 Nov 17;59(4):675–680. doi: 10.1016/0092-8674(89)90013-5. [DOI] [PubMed] [Google Scholar]
  12. Gonzalez G. A., Yamamoto K. K., Fischer W. H., Karr D., Menzel P., Biggs W., 3rd, Vale W. W., Montminy M. R. A cluster of phosphorylation sites on the cyclic AMP-regulated nuclear factor CREB predicted by its sequence. Nature. 1989 Feb 23;337(6209):749–752. doi: 10.1038/337749a0. [DOI] [PubMed] [Google Scholar]
  13. Gottesman M. M., LeCam A., Bukowski M., Pastan I. Isolation of multiple classes of mutants of CHO cells resistant to cyclic AMP. Somatic Cell Genet. 1980 Jan;6(1):45–61. doi: 10.1007/BF01538695. [DOI] [PubMed] [Google Scholar]
  14. Hai T. W., Liu F., Coukos W. J., Green M. R. Transcription factor ATF cDNA clones: an extensive family of leucine zipper proteins able to selectively form DNA-binding heterodimers. Genes Dev. 1989 Dec;3(12B):2083–2090. doi: 10.1101/gad.3.12b.2083. [DOI] [PubMed] [Google Scholar]
  15. Hammond R. A., McClung J. K., Miller M. R. Effect of DNA polymerase inhibitors on DNA repair in intact and permeable human fibroblasts: evidence that DNA polymerases delta and beta are involved in DNA repair synthesis induced by N-methyl-N'-nitro-N-nitrosoguanidine. Biochemistry. 1990 Jan 9;29(1):286–291. doi: 10.1021/bi00453a039. [DOI] [PubMed] [Google Scholar]
  16. Howard P., Day K. H., Kim K. E., Richardson J., Thomas J., Abraham I., Fleischmann R. D., Gottesman M. M., Maurer R. A. Decreased catalytic subunit mRNA levels and altered catalytic subunit mRNA structure in a cAMP-resistant Chinese hamster ovary cell line. J Biol Chem. 1991 Jun 5;266(16):10189–10195. [PubMed] [Google Scholar]
  17. Kedar P. S., Widen S. G., Englander E. W., Fornace A. J., Jr, Wilson S. H. The ATF/CREB transcription factor-binding site in the polymerase beta promoter mediates the positive effect of N-methyl-N'-nitro-N-nitrosoguanidine on transcription. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3729–3733. doi: 10.1073/pnas.88.9.3729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lamph W. W., Dwarki V. J., Ofir R., Montminy M., Verma I. M. Negative and positive regulation by transcription factor cAMP response element-binding protein is modulated by phosphorylation. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4320–4324. doi: 10.1073/pnas.87.11.4320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lin C. S., Goldthwait D. A., Samols D. Induction of transcription from the long terminal repeat of Moloney murine sarcoma provirus by UV-irradiation, x-irradiation, and phorbol ester. Proc Natl Acad Sci U S A. 1990 Jan;87(1):36–40. doi: 10.1073/pnas.87.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maekawa T., Sakura H., Kanei-Ishii C., Sudo T., Yoshimura T., Fujisawa J., Yoshida M., Ishii S. Leucine zipper structure of the protein CRE-BP1 binding to the cyclic AMP response element in brain. EMBO J. 1989 Jul;8(7):2023–2028. doi: 10.1002/j.1460-2075.1989.tb03610.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matsumoto Y., Bogenhagen D. F. Repair of a synthetic abasic site in DNA in a Xenopus laevis oocyte extract. Mol Cell Biol. 1989 Sep;9(9):3750–3757. doi: 10.1128/mcb.9.9.3750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miller M. R., Chinault D. N. The roles of DNA polymerases alpha, beta, and gamma in DNA repair synthesis induced in hamster and human cells by different DNA damaging agents. J Biol Chem. 1982 Sep 10;257(17):10204–10209. [PubMed] [Google Scholar]
  23. Mosbaugh D. W., Linn S. Excision repair and DNA synthesis with a combination of HeLa DNA polymerase beta and DNase V. J Biol Chem. 1983 Jan 10;258(1):108–118. [PubMed] [Google Scholar]
  24. Oertel W. H., Mugnaini E., Tappaz M. L., Weise V. K., Dahl A. L., Schmechel D. E., Kopin I. J. Central GABAergic innervation of neurointermediate pituitary lobe: biochemical and immunocytochemical study in the rat. Proc Natl Acad Sci U S A. 1982 Jan;79(2):675–679. doi: 10.1073/pnas.79.2.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Randahl H., Elliott G. C., Linn S. DNA-repair reactions by purified HeLa DNA polymerases and exonucleases. J Biol Chem. 1988 Sep 5;263(25):12228–12234. [PubMed] [Google Scholar]
  26. Rehfuss R. P., Walton K. M., Loriaux M. M., Goodman R. H. The cAMP-regulated enhancer-binding protein ATF-1 activates transcription in response to cAMP-dependent protein kinase A. J Biol Chem. 1991 Oct 5;266(28):18431–18434. [PubMed] [Google Scholar]
  27. Roesler W. J., Vandenbark G. R., Hanson R. W. Cyclic AMP and the induction of eukaryotic gene transcription. J Biol Chem. 1988 Jul 5;263(19):9063–9066. [PubMed] [Google Scholar]
  28. Siedlecki J. A., Szyszko J., Pietrzykowska I., Zmudzka B. Evidence implying DNA polymerase beta function in excision repair. Nucleic Acids Res. 1980 Jan 25;8(2):361–375. doi: 10.1093/nar/8.2.361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Singh T. J., Hochman J., Verna R., Chapman M., Abraham I., Pastan I. H., Gottesman M. M. Characterization of a cyclic AMP-resistant Chinese hamster ovary cell mutant containing both wild-type and mutant species of type I regulatory subunit of cyclic AMP-dependent protein kinase. J Biol Chem. 1985 Nov 15;260(26):13927–13933. [PubMed] [Google Scholar]
  30. Singh T. J., Roth C., Gottesman M. M., Pastan I. H. Characterization of cyclic AMP-resistant Chinese hamster ovary cell mutants lacking type I protein kinase. J Biol Chem. 1981 Jan 25;256(2):926–932. [PubMed] [Google Scholar]
  31. Smith C. A., Okumoto D. S. Nature of DNA repair synthesis resistant to inhibitors of polymerase alpha in human cells. Biochemistry. 1984 Mar 27;23(7):1383–1391. doi: 10.1021/bi00302a008. [DOI] [PubMed] [Google Scholar]
  32. Stein B., Rahmsdorf H. J., Steffen A., Litfin M., Herrlich P. UV-induced DNA damage is an intermediate step in UV-induced expression of human immunodeficiency virus type 1, collagenase, c-fos, and metallothionein. Mol Cell Biol. 1989 Nov;9(11):5169–5181. doi: 10.1128/mcb.9.11.5169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Teitz T., Penner M., Eli D., Stark M., Bakhanashvili M., Naiman T., Canaani D. Isolation by polymerase chain reaction of a cDNA whose product partially complements the ultraviolet sensitivity of xeroderma pigmentosum group C cells. Gene. 1990 Mar 15;87(2):295–298. doi: 10.1016/0378-1119(90)90316-j. [DOI] [PubMed] [Google Scholar]
  34. Wang T. S., Korn D. Reactivity of KB cell deoxyribonucleic acid polymerases alpha and beta with nicked and gapped deoxyribonucleic acid. Biochemistry. 1980 Apr 29;19(9):1782–1790. doi: 10.1021/bi00550a009. [DOI] [PubMed] [Google Scholar]
  35. Widen S. G., Kedar P., Wilson S. H. Human beta-polymerase gene. Structure of the 5'-flanking region and active promoter. J Biol Chem. 1988 Nov 15;263(32):16992–16998. [PubMed] [Google Scholar]
  36. Widen S. G., Wilson S. H. Mammalian beta-polymerase promoter: large-scale purification and properties of ATF/CREB palindrome binding protein from bovine testes. Biochemistry. 1991 Jun 25;30(25):6296–6305. doi: 10.1021/bi00239a031. [DOI] [PubMed] [Google Scholar]
  37. Wiebauer K., Jiricny J. Mismatch-specific thymine DNA glycosylase and DNA polymerase beta mediate the correction of G.T mispairs in nuclear extracts from human cells. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5842–5845. doi: 10.1073/pnas.87.15.5842. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wilson S., Abbotts J., Widen S. Progress toward molecular biology of DNA polymerase beta. Biochim Biophys Acta. 1988 Feb 28;949(2):149–157. doi: 10.1016/0167-4781(88)90078-4. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES