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Involvement of global genome repair, transcription coupled repair, and chromatin remodeling in UV DNA damage response changes during development.

Lans H, Marteijn JA, Schumacher B, Hoeijmakers JH, Jansen G, Vermeulen W - PLoS Genet. (2010)

Bottom Line: Currently, it is not well understood to what extent both pathways contribute to genome maintenance and cell survival in a developing organism exposed to UV light.Here, we show that eukaryotic NER, initiated by two distinct subpathways, is well conserved in the nematode Caenorhabditis elegans.In germ cells and early embryos, we find that GGR is the major pathway contributing to normal development and survival after UV irradiation, whereas in later developmental stages TCR is predominantly engaged.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Medical Genetics Center, Erasmus MC, Rotterdam, The Netherlands.

ABSTRACT
Nucleotide Excision Repair (NER), which removes a variety of helix-distorting lesions from DNA, is initiated by two distinct DNA damage-sensing mechanisms. Transcription Coupled Repair (TCR) removes damage from the active strand of transcribed genes and depends on the SWI/SNF family protein CSB. Global Genome Repair (GGR) removes damage present elsewhere in the genome and depends on damage recognition by the XPC/RAD23/Centrin2 complex. Currently, it is not well understood to what extent both pathways contribute to genome maintenance and cell survival in a developing organism exposed to UV light. Here, we show that eukaryotic NER, initiated by two distinct subpathways, is well conserved in the nematode Caenorhabditis elegans. In C. elegans, involvement of TCR and GGR in the UV-induced DNA damage response changes during development. In germ cells and early embryos, we find that GGR is the major pathway contributing to normal development and survival after UV irradiation, whereas in later developmental stages TCR is predominantly engaged. Furthermore, we identify four ISWI/Cohesin and four SWI/SNF family chromatin remodeling factors that are implicated in the UV damage response in a developmental stage dependent manner. These in vivo studies strongly suggest that involvement of different repair pathways and chromatin remodeling proteins in UV-induced DNA repair depends on developmental stage of cells.

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Related in: MedlinePlus

Genetic interactions of isw-1 and pbrm-1 with rad-23 and csb-1.(A) L1 larvae and germ cell and embryo UV survival of wild type, rad-23 and csb-1 animals grown on control or isw-1 RNAi food. (B) L1 larvae and germ cell and embryo UV survival of wild type, rad-23, csb-1, pbrm-1, pbrm-1; rad-23 and pbrm-1; csb-1 animals. Each line represents the mean of at least two independent experiments (typically, n>40). Error bars denote the s.e.m.
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pgen-1000941-g008: Genetic interactions of isw-1 and pbrm-1 with rad-23 and csb-1.(A) L1 larvae and germ cell and embryo UV survival of wild type, rad-23 and csb-1 animals grown on control or isw-1 RNAi food. (B) L1 larvae and germ cell and embryo UV survival of wild type, rad-23, csb-1, pbrm-1, pbrm-1; rad-23 and pbrm-1; csb-1 animals. Each line represents the mean of at least two independent experiments (typically, n>40). Error bars denote the s.e.m.

Mentions: The specific UV sensitivity of L1 larvae but not germ cells caused by knockdown of certain chromatin remodeling genes suggests these genes might be involved in TCR but not GGR. If this is the case, knockdown of these genes in a GGR-deficient background could lead to an even more pronounced UV sensitivity, similar as observed for the rad-23; csb-1 double mutants. Likewise, genes that affect UV sensitivity in L1 larvae as well as germ cells might be generally involved in NER, in both TCR and GGR. Inactivation of these genes in a GGR- or TCR-deficient background should not lead to increased UV sensitivity. To test this, we inactivated isw-1, which affects sensitivity in germ cells and L1 larvae, and pbrm-1, which only affects L1 larvae sensitivity, in rad-23 and csb-1 mutants. RNAi-mediated knockdown of isw-1 in rad-23 and csb-1 animals did not lead to significantly enhanced UV sensitivity compared to the respective controls, in both the L1 as well as the germ cell and embryo survival assay (Figure 8A). Only a mild, but reproducible increase in UV sensitivity was observed in the germ cell and embryo sensitivity of rad-23 mutants in which isw-1 was knocked down. Most of these results, however, are in line with the idea that isw-1 has a general regulatory role in the UV-DDR but not specifically in either TCR or GGR.


Involvement of global genome repair, transcription coupled repair, and chromatin remodeling in UV DNA damage response changes during development.

Lans H, Marteijn JA, Schumacher B, Hoeijmakers JH, Jansen G, Vermeulen W - PLoS Genet. (2010)

Genetic interactions of isw-1 and pbrm-1 with rad-23 and csb-1.(A) L1 larvae and germ cell and embryo UV survival of wild type, rad-23 and csb-1 animals grown on control or isw-1 RNAi food. (B) L1 larvae and germ cell and embryo UV survival of wild type, rad-23, csb-1, pbrm-1, pbrm-1; rad-23 and pbrm-1; csb-1 animals. Each line represents the mean of at least two independent experiments (typically, n>40). Error bars denote the s.e.m.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2865526&req=5

pgen-1000941-g008: Genetic interactions of isw-1 and pbrm-1 with rad-23 and csb-1.(A) L1 larvae and germ cell and embryo UV survival of wild type, rad-23 and csb-1 animals grown on control or isw-1 RNAi food. (B) L1 larvae and germ cell and embryo UV survival of wild type, rad-23, csb-1, pbrm-1, pbrm-1; rad-23 and pbrm-1; csb-1 animals. Each line represents the mean of at least two independent experiments (typically, n>40). Error bars denote the s.e.m.
Mentions: The specific UV sensitivity of L1 larvae but not germ cells caused by knockdown of certain chromatin remodeling genes suggests these genes might be involved in TCR but not GGR. If this is the case, knockdown of these genes in a GGR-deficient background could lead to an even more pronounced UV sensitivity, similar as observed for the rad-23; csb-1 double mutants. Likewise, genes that affect UV sensitivity in L1 larvae as well as germ cells might be generally involved in NER, in both TCR and GGR. Inactivation of these genes in a GGR- or TCR-deficient background should not lead to increased UV sensitivity. To test this, we inactivated isw-1, which affects sensitivity in germ cells and L1 larvae, and pbrm-1, which only affects L1 larvae sensitivity, in rad-23 and csb-1 mutants. RNAi-mediated knockdown of isw-1 in rad-23 and csb-1 animals did not lead to significantly enhanced UV sensitivity compared to the respective controls, in both the L1 as well as the germ cell and embryo survival assay (Figure 8A). Only a mild, but reproducible increase in UV sensitivity was observed in the germ cell and embryo sensitivity of rad-23 mutants in which isw-1 was knocked down. Most of these results, however, are in line with the idea that isw-1 has a general regulatory role in the UV-DDR but not specifically in either TCR or GGR.

Bottom Line: Currently, it is not well understood to what extent both pathways contribute to genome maintenance and cell survival in a developing organism exposed to UV light.Here, we show that eukaryotic NER, initiated by two distinct subpathways, is well conserved in the nematode Caenorhabditis elegans.In germ cells and early embryos, we find that GGR is the major pathway contributing to normal development and survival after UV irradiation, whereas in later developmental stages TCR is predominantly engaged.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics, Medical Genetics Center, Erasmus MC, Rotterdam, The Netherlands.

ABSTRACT
Nucleotide Excision Repair (NER), which removes a variety of helix-distorting lesions from DNA, is initiated by two distinct DNA damage-sensing mechanisms. Transcription Coupled Repair (TCR) removes damage from the active strand of transcribed genes and depends on the SWI/SNF family protein CSB. Global Genome Repair (GGR) removes damage present elsewhere in the genome and depends on damage recognition by the XPC/RAD23/Centrin2 complex. Currently, it is not well understood to what extent both pathways contribute to genome maintenance and cell survival in a developing organism exposed to UV light. Here, we show that eukaryotic NER, initiated by two distinct subpathways, is well conserved in the nematode Caenorhabditis elegans. In C. elegans, involvement of TCR and GGR in the UV-induced DNA damage response changes during development. In germ cells and early embryos, we find that GGR is the major pathway contributing to normal development and survival after UV irradiation, whereas in later developmental stages TCR is predominantly engaged. Furthermore, we identify four ISWI/Cohesin and four SWI/SNF family chromatin remodeling factors that are implicated in the UV damage response in a developmental stage dependent manner. These in vivo studies strongly suggest that involvement of different repair pathways and chromatin remodeling proteins in UV-induced DNA repair depends on developmental stage of cells.

Show MeSH
Related in: MedlinePlus