Background: DNA damage leads to cellular responses that include the increased expression of DNA repair genes, repression of DNA replication and alterations in cellular metabolism. Archaeal information processing pathways resemble those in eukaryotes, but archaeal damage response pathways remain poorly understood.

Relatively low concentrations of reactive oxygen cause reversible alterations of endothelial cell signal transduction and gene transcription. The hypothesis that low levels of oxidant stress activate retention of trans-acting proteins in the nucleus was investigated by determining time and dose requirements for oxidant-stimulated nuclear protein binding to consensus DNA sequences for nuclear factor (NF)-kappa B or activator protein 1 (AP-1). Nuclear proteins were extracted from low passage porcine aortic endothelial cells 15 min to 24 h after addition of increasing concentrations of H2O2. Electrophoretic mobility shift assays demonstrated that protein binding to NF-kappa B and AP-1 sequences increases over 1-2 h after stress relative to time-matched controls and resolves by 24 h. The selective protein kinase C inhibitor, calphostin C, prevents approximately 30% of this increase. Inhibition of tyrosine kinase activity by herbimycin A (5 microM) completely inhibits the response to H2O2. Exposure of intact cells to H2O2 increases substrate phosphorylation in pp60src immunoprecipitates. The activity of pp60src in immunoprecipitates from control cells or of recombinant pp60src increases after in vitro addition of H2O2. H2O2-stimulated pp60src activity is reduced by pretreatment of the enzyme preparation with N-acetylcysteine. These data indicate that oxidants increase nuclear levels of trans-acting factors in endothelial cells and that these increases require oxidant-sensitive changes in both tyrosine and serine/threonine phosphorylations.

SummaryThe nucleotide excision repair (NER) pathway removes bulky lesions such as photoproducts from DNA. In both bacteria and eukarya, lesions located in transcribed strands are repaired significantly faster than those located in non-transcribed strands due to damage signalling by stalled RNA polymerase molecules: a phenomenon known as transcriptioncoupled repair (TCR). TCR requires a mechanism for coupling the detection of stalled RNA polymerase molecules to the NER pathway, provided in bacteria by the Mfd protein. In the third domain of life, archaea, the pathway of NER is not well defined, there are no Mfd homologues and the existence of TCR has not been investigated. In this report we looked at rates of removal of photoproducts in three different operons of the crenarchaeon Sulfolobus solfataricus following UV irradiation. We found no evidence for significantly faster repair in the transcribed strands of these three operons. The rate of global genome repair in S. solfataricus is relatively rapid, and this may obviate the requirement for a specialized TCR pathway. Significantly faster repair kinetics were observed in the presence of visible light, consistent with the presence of a gene for photolyase in the genome of S. solfataricus.