Project description:RNA-seq analysis of adult wildtype C.elegans worms which were exposed to two concentrations of H2S (50 and 150ppm) or oxygen control (7% O2), over time (1,2,12h). A likely mixup of some samples was noted based on clustering analysis, and metadata here has been corrected. The original filename contains the initial annotation.
Project description:Compare the global gene expression profiles of Sma-6 and Dbl-1 in L4 stage C. elegans Experiment Overall Design: Sma-6 and Dbl-1 mutants compared
Project description:Vesicle-mediated transport is a fundamental part of the secretory and endocytic pathways. In addition to their role in membrane protein trafficking, Arf-like protein subfamily of small GTPases also plays an important role in development, maintenance of Golgi structure and function. Proper protein trafficking is critical for cellular integrity and studies demonstrate that aberrant protein sorting leads to various diseases in many organisms including humans. However, to date, there is no report showing the role of Golgi apparatus and Golgi proteins in organismal longevity. Organisms dynamically reprogram their transcriptome and proteome as a response to internal and external changes, which alter physiological processes including aging. Compared to many transcriptomic studies that identified aging-regulatory genes, proteomic studies that identified aging-regulatory proteins are relatively scarce. By using a quantitative proteomic approach, we identified MON-2, an Arf-Gef protein implicated in Golgi-to-endosome trafficking, as a longevity-promoting protein. We found that MON-2 is essential for the long lifespan of various longevity mutants. Our results demonstrate that Golgi-to-endosome trafficking is an integral part of lifespan regulation.
Project description:Temperature is one of the primary environmental factors that affect aging, in which protein phosphorylation is an important regulator. Currently, the understanding of phosphorylation events in regulatory networks during aging has remained rather limited. Therefore, the phosphoproteomes of C.elegans of different age groups cultured at 20°C in natural aging process and 25°C in accelerated aging process were analyzed. Through using the iTRAQ-labeled phosphoproteomics method, 2375 phosphoproteins and 9063 phosphosites were identified. Volcano plots illustrated that 208 proteins during natural aging and 130 during accelerated aging, were significantly changed. Gene ontology and pathway analysis revealed that these proteins were mainly involved in temperature response, DNA transcription, protein translation, tissue system development and animal behavior processes. Moreover, our results uncovered those kinases CK2, MAPK and CAMK2 might play important roles in aging regulation. In summary, our results provided a new insight into the complicated phosphor-regulatory network system during aging and an important resource for future studies of protein phosphorylation in worms.
Project description:Our aim was to identify the genome-wide direct LET-418 binding sites in early C.elegans embryos. a FLAG-tagged version of LET-418 was expressed in let-418(ts) mutants synchronised as early mixed stage embryos, and processed for ChIP followed by deep-sequencing. The obtained binding profile was compared to non-enriched input DNA and significant binding sites were identified.
Project description:The response of the nematode C. elegans to Y. pestis infection was evaluated by gene expression profiling. A synchronized population of nematodes were exposed to Y. pestis KIM5 for 24h. Transcript levels from Y. pestis-treated animals were compared with animals maintained on relatively nonpathogenic E. coli OP50 for 24h. Three independent RNA isolations were performed following exposure to either Y. pestis KIM5 or E. coli OP50. Exposures to the different pathogens were performed in parallel for each replicate isolation.
Project description:Transcriptome profiling of Lir3 C.elegans mutants. Larval stage L4 worms were used for RNA isolation from N2 (Bristol), Q40 (polyglutamine model) genetic background; Lir3 and wiltype controls. Experiments were done in triplicates using ribosomal RNA depletion. Libraries were sequenced with 50bp reads on Illumina HiSeq2500 platform
Project description:To provide insights into the mechanism underlying the enhanced immunity of tag-24/octr-1 animals, we used genome microarrays to find clusters of genes commonly misregulated in tag-24 relative to wild-type animals grown on live P. aeruginosa. Gravid adult wild-type and tag-24/octr-1 animals were lysed using a solution of sodium hydroxide and bleach (ratio of 5:2), washed and the eggs were synchronized for 22 hours in S basal liquid medium at room temperature. Synchronized L1 larval animals were placed onto NGM plates seeded with E. coli OP50 and grown at 20°C until the animals had reached the L4 larval stage. Animals were collected and washed with M9 buffer before transferring to NGM plates containing P. aeruginosa PA14 for 4 hours at 25°C. After 4 hours, animals were collected and washed with M9 buffer, and RNA was extracted using TRIzol reagent (Invitrogen). Residual genomic DNA was removed by DNase treatment (Ambion Austin, TX).
Project description:Mechanical stress is a measure of internal resistance exhibited by a body or material when external forces, such as compression, tension, bending, etc. are applied. The study of mechanical stress on health and aging is a continuously growing field, as major changes to the extracellular matrix and cell-to-cell adhesions can result in dramatic changes to tissue stiffness during aging and diseased conditions. For example, during normal aging, many tissues including the ovaries, skin, blood vessels, and heart exhibit increased stiffness, which can result in a significant reduction in function of that organ. As such, numerous model systems have recently emerged to study the impact of mechanical and physical stress on cell and tissue health, including cell-culture conditions with matrigels and other surfaces that alter substrate stiffness and ex vivo tissue models that can apply stress directly to organs like muscle or tendons. Here, we sought to develop a novel method in an in vivo, model organism setting to study the impact of mechanical stress on aging, by increasing substrate stiffness in solid agar medium of C. elegans. To our surprise, we found shockingly limited impact of growth of C. elegans on stiffer substrates, including limited effects on cellular health, gene expression, organismal health, stress resilience, and longevity. Overall, our studies reveal that altering substrate stiffness of growth medium for C. elegans have only mild impact on animal health and longevity; however, these impacts were not nominal and open up important considerations for C. elegans biologists in standardizing agar medium choice for experimental assays.