Microinjected photoreactivating enzymes from Anacystis and Saccharomyces monomerize dimers in chromatin of human cells

Zwetsloot, J.C.M.; Vermeulen, Wim; Hoeijmakers, Jan H.J.; Yasui, Akira; Eker, A.P.M.; Bootsma, D.

doi:10.1016/0167-8817(85)90057-4

Cited by 16 publications

(12 citation statements)

References 37 publications

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“…Yet, we have shown that rapid photoreactivation of CPDs in β‐act‐CPD photolyase transgenic mouse cells has a pronounced effect on NER, as evident from the ∼3‐fold (light‐dependent) reduction in UDS. A comparable reduction in UDS has been observed after microinjection of Anacystis nidulans CPD photolyase into cultured human fibroblasts (Zwetsloot et al ., 1985, 1986), which is somewhat surprising as rodent cells, in contrast to human cells, hardly remove CPD lesions by GG‐NER. Apparently, TC‐NER of CPD lesions constitutes a prominent portion of NER activity in the first hour after UV exposure.…”

Section: Discussionsupporting

confidence: 68%

Enhanced repair of cyclobutane pyrimidine dimers and improved UV resistance in photolyase transgenic mice

et al. 2002

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During evolution, placental mammals appear to have lost cyclobutane pyrimidine dimer (CPD) photolyase, an enzyme that efficiently removes UV-induced CPDs from DNA in a light-dependent manner. As a consequence, they have to rely solely on the more complex, and for this lesion less efficient, nucleotide excision repair pathway. To assess the contribution of poor repair of CPDs to various biological effects of UV, we generated mice expressing a marsupial CPD photolyase transgene. Expression from the ubiquitous beta-actin promoter allowed rapid repair of CPDs in epidermis and dermis. UV-exposed cultured dermal fibroblasts from these mice displayed superior survival when treated with photoreactivating light. Moreover, photoreactivation of CPDs in intact skin dramatically reduced acute UV effects like erythema (sunburn), hyperplasia and apoptosis. Mice expressing the photolyase from keratin 14 promoter photo reactivate CPDs in basal and early differentiating keratinocytes only. Strikingly, in these animals, the anti-apoptotic effect appears to extend to other skin compartments, suggesting the presence of intercellular apoptotic signals. Thus, providing mice with CPD photolyase significantly improves repair and uncovers the biological effects of CPD lesions.

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Section: Discussionsupporting

confidence: 68%

Enhanced repair of cyclobutane pyrimidine dimers and improved UV resistance in photolyase transgenic mice

et al. 2002

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“…In addition, the structure of CPD and (6 -4)PP lesions, their DNA distorting properties, and the interactions between photolysase and DNA has widened the scope for the biological impact of this unique repair enzyme system [87]. After a microinjected microbial photolyase was shown to rapidly eliminate CPD lesions in UVexposed human fibroblasts [88], in the following years several research groups generated mammalian cells transiently or stably expressing photolyases. Experiments with CPD-photolyase transgenes in normal (NER-proficient) human cell lines revealed that CPDs are an instigator of UV-mediated apoptosis [89,90].…”

Section: Photolyase In Mammalsmentioning

confidence: 99%

DNA Repair in Mammalian Cells

Eker

Quayle

Chaves

et al. 2009

Cell. Mol. Life Sci.

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The genomic integrity of all living organisms is constantly jeopardized by physical [e.g. ultraviolet (UV) light, ionizing radiation] and chemical (e.g. environmental pollutants, endogenously produced reactive metabolites) agents that damage the DNA. To overcome the deleterious effects of DNA lesions, nature evolved a number of complex multi-protein repair processes with broad, partially overlapping substrate specificity. In marked contrast, cells may use very simple repair systems, referred to as direct DNA damage reversal, that rely on a single protein, remove lesions in a basically error-free manner, show high substrate specificity, and do not involve incision of the sugar-phosphate backbone or base excision. This concise review deals with two types of direct DNA damage reversal: (i) the repair of alkylating damage by alkyltransferases and dioxygenases, and (ii) the repair of UV-induced damage by spore photoproduct lyases and photolyases. (Part of a Multi-author Review).

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“…Photoreactivation is an elegant reaction to investigate DNA damage recognition in chromatin in living cells, since only one enzyme is involved in this process and its activity can be regulated precisely by light. Photoreactivation experiments with chicken embryo fibroblasts (van de Merwe and Bronk, 1981) and injection of photolyases from yeast and A.nidulans in human fibroblasts (Zwetsloot et al ., 1985) documented that a significant fraction of CPDs can be repaired despite packaging of DNA in chromatin. Another study showed that ∼75% of the DNA was shielded from photorepair immediately after UV exposure, but all sites became available after 9–12 h (Pendrys, 1983).…”

Section: Dna Repair By Photolyase In Chromatinmentioning

confidence: 99%

Light and dark in chromatin repair: repair of UV-induced DNA lesions by photolyase and nucleotide excision repair

1999

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Nucleotide excision repair (NER) and DNA repair by photolyase in the presence of light (photoreactivation) are the major pathways to remove UV-induced DNA lesions from the genome, thereby preventing mutagenesis and cell death. Photoreactivation was found in many prokaryotic and eukaryotic organisms, but not in mammals, while NER seems to be universally distributed. Since packaging of eukaryotic DNA in nucleosomes and higher order chromatin structures affects DNA structure and accessibility, damage formation and repair are coupled intimately to structural and dynamic properties of chromatin. Here, I review recent progress in the study of repair of chromatin and transcribed genes. Photoreactivation and NER are discussed as examples of how an individual enzyme and a complex repair pathway, respectively, access DNA lesions in chromatin and how these two repair processes fulfil complementary roles in removal of UV lesions. These repair pathways provide insight into the structural and dynamic properties of chromatin and suggest how other DNA repair processes could work in chromatin. Keywords: chromatin/DNA repair/nucleotide excision repair/photolyase/transcription Chromatin, a dynamic and heterogeneous substrate for DNA repairChromatin is the packaged state of DNA in the nucleus and the substrate for all DNA-dependent processes, including transcription and DNA repair (Wolffe, 1995). At its lowest level of structural organization, DNA is folded in an array of nucleosomes called the nucleosome filament. The core of a nucleosome is composed of an octamer of four highly folded histone proteins, H2A, H2B, H3 and H4 (two of each), and 145 bp of DNA wrapped around the octamer. The nucleosome filament is formed by connecting nucleosome cores with linker DNA of variable length and by association of one histone H1 per nucleosome. The nucleosome filament is condensed further into compact 30 nm fibres and higher order structures. These structures are present in interphase and metaphase and are the target for damage formation and repair. There is pronounced structural and functional heterogeneity along chromosomal DNA with respect to DNA sequence, composition and functional activity, which projects into heterogeneity of damage formation and repair. This heterogeneity includes © European Molecular Biology Organization 6585 regions required for regulation of gene expression and replication, which associate with sequence-specific proteins and frequently are characterized by enhanced accessibility to nucleases (nuclease-sensitive regions; Workman and Kingston, 1998). Only a minor fraction of genes are transcribed or pre-set for transcription at any moment. The structure of these genes may include histone proteins modified by acetylation, altered or disrupted nucleosomes in the promoter regions, an unfolding of the whole chromatin domain, a loss or rearrangement of nucleosomes in the transcribed part, as well as differential loading of the genes with transcribing RNA polymerases. Moreover, a large fraction of genes are inactivated in h...

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Microinjected photoreactivating enzymes from Anacystis and Saccharomyces monomerize dimers in chromatin of human cells

Cited by 16 publications

References 37 publications

Enhanced repair of cyclobutane pyrimidine dimers and improved UV resistance in photolyase transgenic mice

Enhanced repair of cyclobutane pyrimidine dimers and improved UV resistance in photolyase transgenic mice

DNA Repair in Mammalian Cells

Light and dark in chromatin repair: repair of UV-induced DNA lesions by photolyase and nucleotide excision repair

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