Project description:How epigenetic information is transmitted from generation to generation remains largely unknown. Deletion of the C. elegans Histone H3 lysine 4 dimethyl (H3K4me2) demethylase spr-5 leads to inherited accumulation of the euchromatic H3K4me2 mark and progressive decline in fertility. Here we identified a genetic network of chromatin-modifying factors, including the H3K4me1/me2 methyltransferases SET-17 and SET-30, the H3K9me1/me2 methyltransferase MET-2, an H3K9me3 methyltransferase, SET-26, the H3K9me3 demethylase JMJD-2, and an H3K9me reader EAP-1, which regulate the trans-generational flow of epigenetic information. Importantly, genetic ablation of set-17, set-30, jmjd-2, or eap-1 suppresses the progressive transgenerational phenotypes, while loss of SET-26 or MET-2 accelerates the infertility of spr-5 mutant worms. We further show that loss of spr-5 also causes a trans-generational increase in lifespan, which is dependent on these chromatin regulators as well as DAF-36 and DAF-12, which control a germline to soma longevity signaling pathway. These findings suggest that the balance between the euchromatic H3K4 and the heterochromatic H3K9 methylation regulates trans-generational effects on longevity and fertility. EAP-1 binding ChIPseq libraries were prepared from 50 ul of packed young adult worms which were maintained at 16 degrees until the appropriate generation and then shifted to 25 degrees after birth. Two biological repeats were generated for wildtype and generation 20 spr-5(by101) mutant worms and a single repeat for generation 10. There were four input samples and eap-1 null mutant worms were also analyzed.
Project description:Microarray-based expression profiling of mixed stage populations taken from generation 1,13 and 26 spr-5 mutant animals as well as wild-type animals reveals a large class of spermatogenesis-expressed genes whose expression coordinately increased from generations 1 to 13 and then decreased from generations 13 to 26 in spr-5(by101) mutants. These results suggest that a failure to reset spermatogenesis acquired H3K4me2 may result in the progressive sterility that is observed in spr-5 mutants. 8ug of total RNA was hybridized pair-wise, along with a dye flip, from N2 (wild-type) f1 and spr-5(by101) mutant f1, f13 and f26, for a total of 12 arrays. RNA was labelled with the Genisphere Array350 system.
Project description:Microarray-based expression profiling of mixed stage populations taken from generation 1,13 and 26 spr-5 mutant animals as well as wild-type animals reveals a large class of spermatogenesis-expressed genes whose expression coordinately increased from generations 1 to 13 and then decreased from generations 13 to 26 in spr-5(by101) mutants. These results suggest that a failure to reset spermatogenesis acquired H3K4me2 may result in the progressive sterility that is observed in spr-5 mutants.
Project description:Using distinct mutant strains deficient in key aspects of gene silencing machinery, we provide evidence that neuroprotection appears to stem from dsRNAs that originate from the bacteria themselves, and Systemic RNA Interference Defective (SID) mutants deficient in dsRNA-mediated gene silencing do not allow for neuroprotection from the bacteria. Transcriptomic analysis between worms reared on OP50 and HB101 E. coli for multiple generations in an a-syn background revealed that the primary cellular processes involved in neuroprotection of this nature included but were not limited to endopeptidase activity, transcriptional regulation, calcium ion binding activity, and water channel activity.