PgmNr W452: Beyond Cell Death: Systematic Analyses of Non-apoptotic CED-3 Caspase Functions in C. elegans.

Authors:
B. P. Weaver; Y. M. Weaver; M. Han


Institutes
HHMI & CU Boulder, Boulder, USA.


Keyword: Timing of developmental events, and circadian rhythms

Abstract:

Recent findings from several labs, including ours, have implicated important non-apoptotic functions for canonical apoptotic regulators across diverse metazoans including nematodes, flies, and mammals. We recently reported a genome-wide RNAi screen in C. elegans that identified numerous genes cooperating with the miRISC to ensure robust development (Weaver, et al. 2014). We showed that one interactor, the CED-3 caspase, previously known for its essential role in apoptosis, had a critical non-apoptotic activity negatively regulating the expression of LIN-28, and likely also LIN-14 and DISL-2 (Dis3l2 ribonuclease) in the LIN-28 pluripotency/ developmental timing pathway in late larval development thereby limiting supernumerary seam cell divisions (Weaver, et al. 2014). In order to systematically investigate the extent of possible other non-apoptotic functions for the CED-3 caspase in C. elegans development, we performed a ced-3(lf) enhancer screen in the double blind using a genome-wide RNAi approach. Following a secondary RNAi confirmation screen, we identified more than 100 interactors from diverse functional categories including protein stability regulators, signal transduction factors, RNA-binding proteins, and transcriptional regulators, thus implicating the involvement of CED-3 caspase in a wide-range of non-apoptotic developmental regulatory pathways. We sought to identify a downstream target for one of these interactions that resulted in a dramatic larval developmental stall. Using a combination of biochemical methods, genetic methods, and tissue-specific reporters, we found that CED-3 negatively regulated a MAPK pathway in the hypodermis (epidermis), in the absence of stress. We found that aberrant activity of the MAPK pathway was responsible for larval delay and that CED-3 caspase contributed to robust late larval development by downregulating this pathway and thereby ensured proper post-embryonic growth rate during normal development. Altogether, our data support a model wherein CED-3 caspase acts as a pro-differentiation factor in later larval stages that is important to ensure both proper developmental timing (previous findings) while also ensuring timely progress through post-embryonic development (current findings).



Wormbase Genetic Index
1. ced-3
2. lin-28
3. lin-14
4. disl-2