Wang S et al. (2022) Nature "Inheritance of paternal DNA damage by histone-mediated repair restriction."

Meyer DH, Schumacher B, Wang S

[Nature, 2022]

How paternal exposure to ionizing radiation affects genetic inheritance and disease risk in the offspring has been a long-standing question in radiation biology. In humans, nearly 80% of transmitted mutations arise in the paternal germline¹, but the transgenerational effects of ionizing radiation exposure has remained controversial and the mechanisms are unknown. Here we show that in sex-separated Caenorhabditis elegans strains, paternal, but not maternal, exposure to ionizing radiation leads to transgenerational embryonic lethality. The offspring of irradiated males displayed various genome instability phenotypes, including DNA fragmentation, chromosomal rearrangement and aneuploidy. Paternal DNA double strand breaks were repaired by maternally provided error-prone polymerase theta-mediated end joining. Mechanistically, we show that depletion of an orthologue of human histone H1.0, HIS-24, or the heterochromatin protein HPL-1, could significantly reverse the transgenerational embryonic lethality. Removal of HIS-24 or HPL-1 reduced histone 3 lysine 9 dimethylation and enabled error-free homologous recombination repair in the germline of the F₁ generation from ionizing radiation-treated P₀ males, consequently improving the viability of the F₂ generation. This work establishes the mechanistic underpinnings of the heritable consequences of paternal radiation exposure on the health of offspring, which may lead to congenital disorders and cancer in humans.

Wang S et al. (2017) Development "A toolkit for GFP-mediated tissue-specific protein degradation in C. elegans."

Zhao Z, Prevo B, Tang NH, Desai A, Wang S, Cheerambathur DK, Lara-Gonzalez P, Chisholm AD, Oegema K

[Development, 2017]

Proteins essential for embryo production, cell division, and early embryonic events are frequently re-utilized later in embryogenesis, during organismal development, or in the adult. Examining protein function across these different biological contexts requires tissue-specific perturbation. Here, we describe a method that utilizes expression of a fusion between a GFP-targeting nanobody and a SOCS-box containing ubiquitin ligase adaptor to target GFP tagged proteins for degradation. When combined with endogenous locus GFP tagging by CRISPR-Cas9 or rescue of a null mutant with a GFP fusion, this approach enables routine and efficient tissue-specific protein ablation. We show that this approach works in multiple tissues-the epidermis, intestine, body wall muscle, ciliated sensory neurons, and touch receptor neurons-where it recapitulates expected loss-of-function mutant phenotypes. The transgene toolkit and the strain set described here will complement existing approaches to enable routine analysis of the tissue-specific roles of C. elegans proteins.

Wang S et al. (2021) Chemosphere "Induction of protective response to polystyrene nanoparticles associated with methylation regulation in Caenorhabditis elegans."

Zhang R, Wang D, Wang S

[Chemosphere, 2021]

The epigenetic regulation mechanisms for toxicity induction of nanoplastics in organisms remain largely unknown. In Caenorhabditis elegans, we found that prolonged exposure to 1-100g/L polystyrene nanoparticles (PS-NPs) decreased expression of MET-2, a H3K9 methyltransferase. Meanwhile, RNAi knockdown of met-2 suppressed the PS-NPs toxicity in inducing production of reactive oxygen species (ROS) and in decreasing locomotion behavior, which suggesting that the decrease in MET-2 expression reflected a protective response. This resistance to PS-NPs toxicity could be further detected in worms with met-2 RNAi knockdown in both intestinal cells and germline cells. In PS-NPs exposed worms, intestinal RNAi knockdown of met-2 significantly increased expressions of daf-16, bar-1, and elt-2. Intestinal RNAi knockdown of daf-16, bar-1, or elt-2 suppressed the resistance of met-2(RNAi) worms to PS-NPs toxicity, suggesting that MET-2 functioned upstream of ELT-2, BAR-1, and DAF-16 in intestinal cells to control PS-NPs toxicity. Moreover, in PS-NPs exposed worms, germline RNAi knockdown of met-2 significantly decreased expressions of wrt-3 and pat-12. RNAi knockdown of wrt-3 or pat-12 further inhibited the susceptibility of worms overexpressing germline MET-2 to PS-NPs toxicity, suggesting that MET-2 functioned upstream of PAT-12 and WRT-3 in germline cells to control PS-NPs toxicity. Therefore, our data provided an important molecular basis for MET-2-mediated methylation regulation in causing protective response to nanoplastics in organisms.

Wang S et al. (2001) EMBO J "The TRA-1 transcription factor binds TRA-2 to regulate sexual fates in Caenorhabditis elegans."

Wang S, Kimble J

[EMBO J, 2001]

The tra-1 and tra-2 sex-determining genes promote female fates in Caenorhabditis elegans. Classical genetic studies placed tra-1 as the terminal regulator of the pathway with tra-2 acting upstream as a regulator of regulators of tra-1. Here we report the surprising result that the TRA-1 transcription factor binds the intracellular domain of the TRA-2 membrane protein. This binding is dependent on the MX regulatory domain, a region of the TRA-2 intracellular domain shown previously to be critical for the onset of hermaphrodite spermatogenesis. The functional importance of the TRA-1-TRA-2 physical interaction is supported by genetic interactions between tra-1(0) and tra-2(mx) mutations: a reduction of tra-1 gene dose from two copies to one copy enhances the tra-2(mx) feminization phenotype, but has no apparent somatic effect. In Caenorhabditis briggsae, we also find an MX-dependent interaction between Cb-TRA-1 and Cb-TRA-2, hut intriguingly, no cross-species interactions are seen. The conservation of the TRA-1-TRA-2 interaction underscores its importance in sex determination.

Wang S et al. (2020) Nat Struct Mol Biol "H3K4me2 regulates the recovery of protein biosynthesis and homeostasis following DNA damage."

Schumacher B, Wang S, Meyer DH

[Nat Struct Mol Biol, 2020]

DNA damage causes cancer, impairs development and accelerates aging. Transcription-blocking lesions and transcription-coupled repair defects lead to developmental failure and premature aging in humans. Following DNA repair, homeostatic processes need to be reestablished to ensure development and maintain tissue functionality. Here, we report that, in Caenorhabditis elegans, removal of the WRAD complex of the MLL/COMPASS H3K4 methyltransferase exacerbates developmental growth retardation and accelerates aging, while depletion of the H3K4 demethylases SPR-5 and AMX-1 promotes developmental growth and extends lifespan amid ultraviolet-induced damage. We demonstrate that DNA-damage-induced H3K4me2 is associated with the activation of genes regulating RNA transport, splicing, ribosome biogenesis and protein homeostasis and regulates the recovery of protein biosynthesis that ensures survival following genotoxic stress. Our study uncovers a role for H3K4me2 in coordinating the recovery of protein biosynthesis and homeostasis required for developmental growth and longevity after DNA damage.

Wang S et al. (2018) Chemosphere "Cadmium-induced serotonergic neuron and reproduction damages conferred lethality in the nematode Caenorhabditis elegans."

Zhang K, Chu Z, Miao G, Wang S

[Chemosphere, 2018]

Cadmium is a ubiquitous environmental toxicant. The use of Caenorhabditis elegans as a model for monitoring cadmium exposure has revealed several conserved signaling pathways. However, little is known about the killing process during lethality assay. In the present study, we investigated the effects serotonergic neuronal and reproductive damages on cadmium exposure in C.elegans. We found that sterile hermaphrodites, males and worms that passed reproduction span presented high cadmium resistance compared to those of young adults. The results demonstrated that reproduction process other than reproduction capacity conferred cadmium sensitivity. Cadmium exposure resulted in high ratio bagging phenotype, which was a severe reproductive deficit with embryos hatched internally that could cause worms to die early. The mechanism of bagging formation was ascribed to cadmium-induced egg laying deficiency that led embryos to retain and hatch in uterus. The addition of serotonin and imipramine promoted egg laying and thereby increased cadmium resistance. The results demonstrated that vulval muscles responsible for egg laying were still functional, while the serotonergic hermaphrodite specific neurons might be dysfunctional under cadmium exposure. Cadmium exposure resulted in shrinkage of serotonergic neuronal body and reduced expressions of tryptophan hydroxylase, the key enzyme for serotonin synthesis. The protection of serotonergic neuron through transient thermal preconditioning improved survival rate. In conclusion, our study demonstrated that damages of serotonergic neurons and reproduction conferred to cadmium-induced lethality.

Wang S et al. (2021) Ecotoxicol Environ Saf "Response of tyramine and glutamate related signals to nanoplastic exposure in Caenorhabditis elegans."

Wang S, Qu M, Wang D, Liu H

[Ecotoxicol Environ Saf, 2021]

Neurotransmission related signals are involved in the control of response to toxicants. We here focused on the tyramine and the glutamate related signals to determine their roles in regulating nanoplastic toxicity in Caenorhabditis elegans. In the range of g/L, exposure to nanopolystyrene (100nm) increased the expression of tdc-1 encoding a tyrosine decarboxylase required for synthesis of tyramine, and decreased the expression of eat-4 encoding a glutamate transporter. Both TDC-1 and EAT-4 could act in the neurons to regulate the nanopolystyrene toxicity. Meanwhile, neuronal RNAi knockdown of tdc-1 induced a susceptibility to nanopolystyrene toxicity, and neuronal RNAi knockdown of eat-4 induced a resistance to nanopolystyrene toxicity. In the neurons, TYRA-2 functioned as the corresponding receptor of tyramine and acted upstream of MPK-1 signaling to regulate the nanopolystyrene toxicity. Moreover, during the control of nanopolystyrene toxicity, GLR-4 and GLR-8 were identified as the corresponding glutamate receptors, and acted upstream of JNK-1 signaling and DBL-1 signaling, respectively. Our results demonstrated the crucial roles of tyramine and glutamate related signals in regulating the toxicity of nanoplastics in organisms.

Wang S et al. (2019) Development "A high-content imaging approach to profile C. elegans embryonic development."

Ochoa SD, Green RA, Desai A, Hendel JM, Wang S, Oegema K, Zhao Z, Gerson-Gurwitz A, Chisholm AD, Biggs R, Khaliullin RN

[Development, 2019]

The <i>C. elegans</i> embryo is an important model for analyzing mechanisms of cell fate specification and tissue morphogenesis. Sophisticated lineaging approaches for analyzing embryogenesis have been developed but are labor-intensive and do not naturally integrate morphogenetic readouts. To enable the rapid classification of developmental phenotypes, we developed a high-content method that employs two custom strains: a Germ Layer strain expressing nuclear markers in the ectoderm, mesoderm and endoderm/pharynx, and a Morphogenesis strain expressing markers labeling epidermal cell junctions and the neuronal cell surface. We describe a procedure that allows simultaneous live imaging of development in 80-100 embryos and provide a custom program that generates cropped, oriented image stacks of individual embryos to facilitate analysis. We demonstrate the utility of our method by perturbing 40 previously characterized developmental genes in variants of the two strains containing RNAi-sensitizing mutations. The resulting datasets yielded distinct, reproducible signature phenotypes for a broad spectrum of genes involved in cell fate specification and morphogenesis. Our analysis additionally provides new <i>in vivo</i> evidence for MBK-2 function in mesoderm fate specification and LET-381 function in elongation.

Wang S et al. (2011) Dev Biol "Lineage specific trimethylation of H3 on lysine 4 during C. elegans early embryogenesis."

Poulin GB, Wang S, Fisher K

[Dev Biol, 2011]

In many organisms early embryogenesis is characterised by a period refractory to transcription. In Caenorhabditis elegans, the one-cell embryo is transcriptionally inactive, but at around eight-cell stage transcription is activated in the somatic lineage. This model suggests that histone tail modifications associated with activation of transcription, such as di- or trimethylation of histone 3 on lysine 4 (H3K4me2/me3) should be enriched in the somatic lineage. Here, we have investigated the deposition of H3K4me3 during embryogenesis and found that it is more dynamic than anticipated. In the eight-cell stage embryo, H3K4me3 deposition is poor in the germline blastomere, as expected, but surprisingly three somatic blastomeres also remain poor in H3K4me3. All the other somatic blastomeres show robust deposition of H3K4me3. Interestingly, the three somatic blastomeres poor in H3K4me3 are descendants of the first germline blastomere, implying an activity that impedes on H3K4me3 deposition in these cells. In contrast, the deposition of H3K4me2 and H3K27me2/3 is not lineage restricted. Taken together, our data reveal that H3K4me3 deposition is highly regulated according to the cell lineage involved.

Wang S et al. (2007) Chem Res Toxicol "Induction of germline cell cycle arrest and apoptosis by sodium arsenite in Caenorhabditis elegans."

Hei TK, Su C, Wang S, Yu Z, Tang M, Wu L, Zhao Y

[Chem Res Toxicol, 2007]

The nematode Caenorhabditis elegans has been shown to be a model organism in studying aquatic toxicity. Although epidemiological studies have shown that arsenic is teratogenic and carcinogenic to humans, the lethality assay indicated that C. elegans is less sensitive to inorganic arsenic than any other organisms that have been tested thus far. In the present study, we used the more malleable germline of C. elegans as an in vivo system to investigate the genotoxic effects of arsenite. After animals were exposed to sodium arsenite at concentrations ranging from 1 muM to 0.5 mM, mitotic germ cells and germline apoptosis were scored after DAPI staining and acridine orange vital staining, respectively. DMSO rescue experiments were performed by exposing C. elegans to 0.01 mM arsenite in the presence of DMSO (0.1%) for 24 h, and reactive oxygen species (ROS) were semiquantified by CM-H2DCFDA vital staining. The results indicated that arsenic exposure reduced the brood size of C. elegans and caused mitotic cell cycle arrest and germline apoptosis, which, to some extent, exhibited a concentration- and time-dependent manner. The addition of 0.1% DMSO completely rescued arsenic-induced cell cycle arrest and partially suppressed germline apoptosis. Furthermore, treatment of animals with arsenite at a dose of 0.01 mM significantly increased ROS production in the intestine, which could be reduced by DMSO treatment. The present study also indicated that C. elegans might be used as an in vivo model system to study the mechanisms of arsenic-induced genotoxic effects.