Ultraviolet mutagenesis of radiation-sensitive (rad) mutants of the nematode Caenorhabditis elegans

doi:10.1016/0165-7992(88)90025-5

Mutation Research Letters

Volume 209, Issues 3–4, November–December 1988, Pages 99-106

https://doi.org/10.1016/0165-7992(88)90025-5 Get rights and content

Abstract

A mutational tester strain (JP10) of the nematode C. elegans was used to capture recessive lethal mutations in a balanced 300 essential gene autosomal region. The probability of converting a radiation interaction into a lethal mutation was measured in young gravid adults after exposure to fluences of 254-nm ultraviolet radiation (UV) ranging from 0 to 300 Jm⁻². Mutation frequencies as high as 5% were observed. In addition, three different radiation-hypersensitive mutations, rad-1, rad-3 and rad-7 were incorporated into the JP10 background genotype, which allowed us to measure mutation frequencies in radiation-sensitive animals. The strain homozygous for rad-3 was hypermutable to UV while strains homozygous for rad-1 and rad-7 were hypomutable. Data showing the effects of UV on larval development and fertility for the rad mutants is also shown and compared for wild-type and JP10 backgrounds.

References (25)

A.S. Goldman et al.
The effects of monochromatic ultraviolet light on the egg of Drosophila
Exp. Cell Res.
(1956)
P.S. Hartman
Effects of age and liquid holding on the UV-radiation sensitivities of wild-type and mutant Caenorhabditis elegans dauer larvae
Mutation Res.
(1984)
R. Rosenbluth et al.
Mutagenesis in Caenorhabditis elegans
C.F. Arlett et al.
Variation in response to mutagens amongst normal and repair-defective human cells
Basic Life Sci.
(1983)
S. Brenner
The genetics of Caenorhabditis elegans
Genetics
(1974)
T.P. Coohill
Virus-cell interactions as probes for vacuum-ultraviolet radiation damage and repair
Photochemistry and Photobiology
(1986)
E. Friedberg
I.S. Greenwald et al.
Unc-93 (e1500): A behavioral mutant of Caenorhabditis elegans that defines a gene with a wild-type null phenotype
Genetics
(1980)
W. Harm
P.S. Hartman
UV radiation of wild type and radiation sensitive mutants of the nematode Caenorhabditis elegans: fertilities, survival, and parental effects
Photochem. Photobiol.
(1984)

P.S. Hartman

Epistatic interactions of radiationsensitive (rad) mutants of Caenorhabditis elegans

Genetics

(1985)

P.S. Hartman et al.

Radiation sensitive mutants of Caenorhabditis elegans

Genetics

(1982)

Cited by (16)

Comparative phototoxicity of nanoparticulate and bulk ZnO to a free-living nematode Caenorhabditis elegans: The importance of illumination mode and primary particle size
2011, Environmental Pollution
Citation Excerpt :
Another possibility is that nanoparticles were ingested by C. elegans and the toxicity occurred internally. C. elegans unselectively ingest bacteria and fine particles that are less than 5 μm in size (Donkin and Dusenbery, 1993), and its transparency and small body diameter allow for adequate UV penetration (Coohill et al., 1988; Mills and Hartman, 1998). A measure of oxidative stress (e.g., lipid peroxidation) within the nematodes may help to differentiate these two modes of action.
The present study evaluated phototoxicity of nanoparticulate ZnO and bulk-ZnO under natural sunlight (NSL) versus ambient artificial laboratory light (AALL) illumination to a free-living nematode Caenorhabditis elegans. Phototoxicity of nano-ZnO and bulk-ZnO was largely dependent on illumination method as 2-h exposure under NSL caused significantly greater mortality in C. elegans than under AALL. This phototoxicity was closely related to photocatalytic reactive oxygen species (ROS) generation by the ZnO particles as indicated by concomitant methylene blue photodegradation. Both materials caused mortality in C. elegans under AALL during 24-h exposure although neither degraded methylene blue, suggesting mechanisms of toxicity other than photocatalytic ROS generation were involved. Particle dissolution of ZnO did not appear to play an important role in the toxicity observed in this study. Nano-ZnO showed greater phototoxicity than bulk-ZnO despite their similar size of aggregates, suggesting primary particle size is more important than aggregate size in determining phototoxicity.
Nucleotide excision repair genes are expressed at low levels and are not detectably inducible in Caenorhabditis elegans somatic tissues, but their function is required for normal adult life after UVC exposure
2010, Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
We performed experiments to characterize the inducibility of nucleotide excision repair (NER) in Caenorhabditis elegans, and to examine global gene expression in NER-deficient and -proficient strains as well as germline vs. somatic tissues, with and without genotoxic stress. We also carried out experiments to elucidate the importance of NER in the adult life of C. elegans under genotoxin-stressed and control conditions. Adult lifespan was not detectably different between wild-type and NER-deficient xpa-1 nematodes under control conditions. However, exposure to 6 J/m²/day of ultraviolet C radiation (UVC) decreased lifespan in xpa-1 nematodes more than a dose of 100 J/m²/day in wild-type. Similar differential sensitivities were observed for adult size and feeding. Remarkably, global gene expression was nearly identical in young adult wild-type and xpa-1 nematodes, both in control conditions and 3 h after exposure to 50 J/m² UVC. Neither NER genes nor repair activity were detectably inducible in young adults that lacked germ cells and developing embryos (glp-1 strain). However, expression levels of dozens of NER and other DNA damage response genes were much (5–30-fold) lower in adults lacking germ cells and developing embryos, suggesting that somatic and post-mitotic cells have a much lower DNA repair ability. Finally, we describe a refinement of our DNA damage assay that allows damage measurement in single nematodes.
Nucleotide excision repair and the degradation of RNA pol II by the Caenorhabditis elegans XPA and Rsp5 orthologues, RAD-3 and WWP-1
2008, DNA Repair
The Caenorhabditis elegans rad-3 gene was identified in a genetic screen for radiation sensitive (rad) mutants. Here, we report that the UV sensitivity of rad-3 mutants is caused by a nonsense mutation in the C. elegans orthologue of the human nucleotide excision repair gene XPA. We have used the xpa-1/rad-3 mutant to examine how a defect in nucleotide excision repair (NER) perturbs development. We find that C. elegans carrying a mutation in xpa-1/rad-3 are hypersensitive and hypermutable in response to UV irradiation, but do not display hypersensitivity to oxidative stress or show obvious developmental abnormalities in the absence of UV exposure. Consistent with these observations, non-irradiated xpa-1 mutants have a similar lifespan as wild type. We further show that UV irradiated xpa-1 mutants undergo a stage-dependent decline in growth and survival, which is associated with a loss in transcriptional competence. Surprisingly, transcriptionally quiescent dauer stage larvae are able to survive a dose of UV irradiation, which is otherwise lethal to early stage larvae. We show that the loss of transcriptional competence in UV irradiated xpa-1 mutants is associated with the degradation of the large RNA polymerase II (RNA pol II) subunit, AMA-1, and have identified WWP-1 as the putative E3 ubiquitin ligase mediating this process. The absence of wwp-1 by itself does not cause sensitivity to UV irradiation, but it acts synergistically with a mutation in xpa-1 to enhance UV hypersensitivity.
Radiobiological studies with the nematode Caenorhabditis elegans. Genetic and developmental effects of high LET radiation
1992, International Journal of Radiation Applications and Instrumentation. Part
The biological effects of heavy charged particle (HZE) radiation are of particular interest to travellers and planners for long-duration space flights where exposure levels represent a potential health hazard. The unique feature of HZE radiation is the structured pattern of its energy deposition in targets. There are many consequences of this feature to biological endpoints when compared with effects of ionizing photons. Dose vs response and dose-rate kinetics may be modified, DNA and cellular repair systems may be altered in their abilities to cope with damage, and the qualitative features of damage may be unique for different ions. The nematode Caenorhabditis elegans is being used to address these and related questions associated with exposure to radiation. HZE-induced mutation, chromosome aberration, cell inactivation and altered organogenesis are discussed along with plans for radiobiological experiments in space.
Does trans-lesion synthesis explain the UV-radiation resistance of DNA synthesis in C. elegans embryos?
1991, Mutation Research-DNA Repair
Over 10-fold larger fluences were required to inhibit both DNA synthesis and cell divisionin wild-type C. elegans embryos as compared with other model systems or C. elegans rad mutants. In addition, unlike in other organisms, the molecular weight of daughter DNA strands was reduced only after large, superlethal fluences. The molecular weight of nascent DNA fragments exceeded the interdimer distance by up to 19-fold, indicating that C. elegans embryos can replicate through non-instructional lesions. This putative trans-lesion synthetic capability may explain the refractory nature of UV radiation on embryonic DNA synthesis and nuclear division C. elegans.
Most ultraviolet irradiation induced mutations in the nematode Caenorhabditis elegans are chromosomal rearrangements
1991, Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
In this study we have determined the utility of 254-nm ultraviolet light (UV) as a mutagenic tool in C. elegans. We have demonstrated that irradiation of adult hermaphrodites provides a simple method for the induction of heritable chromosomal rearrangements. A screening protocol was employed that identifies either recessive lethal mutations in the 40 map unit region balanced by the translocation eT1(III;V), or unc-36(III) duplications. Mutations were recovered in 3% of the chromosomes screened after a dose of 120 J/m². This rate resembles that for 1500 R γ-ray-induced mutations selected in a similar manner. The mutations were classifie either as lethals [mapping to Linkage Group (LG)III or LGV] or as putative unc-36 duplications. In contrast to the majority of UV-induced mutations analysed in microorganisms, we found that a large fraction of the C. elegans UV-induced mutations are not simple intragenic lesions, but are deficiencies for more than one adjacent gene or more complex events. Preliminary evidence for this conclusion came from the high frequency of mutations that had a dominant effect causing reduced numbers of adult progeny. Subsequently 6 out of 9 analysed LGV mutations were found to be deficiencies. Other specific rearrangements also identified were: one translocation, sT5(II;III), and two unc-36 duplications, sDp8 and sDp9. It was concluded that UV irradiation can easily be used as an additional tool for the analysis of C. elegans chromosomes, and that C. elegans should prove to be a useful organism in which to study the mechanisms whereby UV acts as a mutagen in cells of complex eukaryotes.

View all citing articles on Scopus

View full text

Article preview

Mutation Research Letters

Abstract

References (25)

The effects of monochromatic ultraviolet light on the egg of Drosophila

Exp. Cell Res.

Effects of age and liquid holding on the UV-radiation sensitivities of wild-type and mutant Caenorhabditis elegans dauer larvae

Mutation Res.

Mutagenesis in Caenorhabditis elegans

Variation in response to mutagens amongst normal and repair-defective human cells

Basic Life Sci.

The genetics of Caenorhabditis elegans

Genetics

Virus-cell interactions as probes for vacuum-ultraviolet radiation damage and repair

Photochemistry and Photobiology

Unc-93 (e1500): A behavioral mutant of Caenorhabditis elegans that defines a gene with a wild-type null phenotype

Genetics

UV radiation of wild type and radiation sensitive mutants of the nematode Caenorhabditis elegans: fertilities, survival, and parental effects

Photochem. Photobiol.

Epistatic interactions of radiationsensitive (rad) mutants of Caenorhabditis elegans

Genetics

Radiation sensitive mutants of Caenorhabditis elegans

Genetics

Cited by (16)

Comparative phototoxicity of nanoparticulate and bulk ZnO to a free-living nematode Caenorhabditis elegans: The importance of illumination mode and primary particle size

Nucleotide excision repair genes are expressed at low levels and are not detectably inducible in Caenorhabditis elegans somatic tissues, but their function is required for normal adult life after UVC exposure

Nucleotide excision repair and the degradation of RNA pol II by the Caenorhabditis elegans XPA and Rsp5 orthologues, RAD-3 and WWP-1

Radiobiological studies with the nematode Caenorhabditis elegans. Genetic and developmental effects of high LET radiation

Does trans-lesion synthesis explain the UV-radiation resistance of DNA synthesis in C. elegans embryos?

Most ultraviolet irradiation induced mutations in the nematode Caenorhabditis elegans are chromosomal rearrangements

Research letterUltraviolet mutagenesis of radiation-sensitive (rad) mutants of the nematode Caenorhabditis elegans

Abstract

Exp. Cell Res.

Mutation Res.

Variation in response to mutagens amongst normal and repair-defective human cells

Basic Life Sci.

The genetics of Caenorhabditis elegans

Genetics

Virus-cell interactions as probes for vacuum-ultraviolet radiation damage and repair

Photochemistry and Photobiology

Unc-93 (e1500): A behavioral mutant of Caenorhabditis elegans that defines a gene with a wild-type null phenotype

Genetics

UV radiation of wild type and radiation sensitive mutants of the nematode Caenorhabditis elegans: fertilities, survival, and parental effects

Photochem. Photobiol.

Epistatic interactions of radiationsensitive (rad) mutants of Caenorhabditis elegans

Genetics

Radiation sensitive mutants of Caenorhabditis elegans

Genetics

Research letter
Ultraviolet mutagenesis of radiation-sensitive (rad) mutants of the nematode Caenorhabditis elegans