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

Ella Englander, Samuel H. Wilson

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

DNA β-polymerase (β-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 β-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 β-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 β-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 β-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 β-pol promoter. Thus, these effectors may act through a common pathway. These results suggest that the activation of the cloned β-pol promoter in CHO cells following MNNG treatment is mediated through the cAMP/protein kinase A signal transduction pathway.

Original languageEnglish (US)
Pages (from-to)5527-5531
Number of pages5
JournalNucleic Acids Research
Volume20
Issue number21
StatePublished - Nov 11 1992
Externally publishedYes

Fingerprint

Protein Kinase
CHO Cells
Methylnitronitrosoguanidine
DNA-Directed DNA Polymerase
Cyclic AMP-Dependent Protein Kinases
Promoter
Mutant
DNA Damage
DNA
Damage
Cells
Proteins
Cell Line
Line
Cell
Chemical activation
Activation
Palindrome
DNA Repair
Pathway

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

DNA damage response of cloned DNA β-polymerase promoter is blocked in mutant cell lines deficient in protein kinase A. / Englander, Ella; Wilson, Samuel H.

In: Nucleic Acids Research, Vol. 20, No. 21, 11.11.1992, p. 5527-5531.

Research output: Contribution to journalArticle

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