The unfolded protein response (UPR), a signalling pathway activated by the accumulation of unfolded proteins in the endoplasmic reticulum, is known to contribute to the defence against infectious agents and toxins. Here it is shown that activation of innate immunity in Caenorhabditis elegans induces the UPR mediated by X-box binding protein 1 (XBP-1). This points to an ancient, conserved role for the XBP-1-dependent UPR in protecting cells against the endoplasmic reticulum stress generated from an immune response. The unfolded protein response, known to contribute to the defence against infectious agents and toxins, is shown here to protect Caenorhabditis elegans larvae against detrimental effects of the innate immune response to infection with Pseudomonas aeruginosa. The findings establish innate immunity as a physiologically relevant inducer of ER stress during C. elegans development. The detection and compensatory response to the accumulation of unfolded proteins in the endoplasmic reticulum (ER), termed the unfolded protein response (UPR), represents a conserved cellular homeostatic mechanism with important roles in normal development and in the pathogenesis of disease1. The IRE1–XBP1/Hac1 pathway is a major branch of the UPR that has been conserved from yeast to human2,3,4,5,6. X-box binding protein 1 (XBP1) is required for the differentiation of the highly secretory plasma cells of the mammalian adaptive immune system7,8, but recent work also points to reciprocal interactions between the UPR and other aspects of immunity and inflammation9,10,11. We have been studying innate immunity in the nematode Caenorhabditis elegans, having established a principal role for a conserved PMK-1 p38 mitogen-activated protein kinase (MAPK) pathway in mediating resistance to microbial pathogens12. Here we show that during C. elegans development, XBP-1 has an essential role in protecting the host during activation of innate immunity. Activation of the PMK-1-mediated response to infection with Pseudomonas aeruginosa induces the XBP-1-dependent UPR. Whereas a loss-of-function xbp-1 mutant develops normally in the presence of relatively non-pathogenic bacteria, infection of the xbp-1 mutant with P. aeruginosa leads to disruption of ER morphology and larval lethality. Unexpectedly, the larval lethality phenotype on pathogenic P. aeruginosa is suppressed by loss of PMK-1-mediated immunity. Furthermore, hyperactivation of PMK-1 causes larval lethality in the xbp-1 mutant even in the absence of pathogenic bacteria. Our data establish innate immunity as a physiologically relevant inducer of ER stress during C. elegans development and indicate that an ancient, conserved role for XBP-1 may be to protect the host organism from the detrimental effects of mounting an innate immune response to microbes.

Innate immunity in Caenorhabditis elegans requires a conserved PMK-1 p38 mitogen-activated protein kinase (MAPK) pathway that regulates the basal and pathogen-induced expression of immune effectors. The mechanisms by which PMK-1 p38 MAPK regulates the transcriptional activation of the C. elegans immune response have not been identified. Furthermore, in mammalian systems the genetic analysis of physiological targets of p38 MAPK in immunity has been limited. Here, we show that C. elegans ATF-7, a member of the conserved cyclic AMP–responsive element binding (CREB)/activating transcription factor (ATF) family of basic-region leucine zipper (bZIP) transcription factors and an ortholog of mammalian ATF2/ATF7, has a pivotal role in the regulation of PMK-1–mediated innate immunity. Genetic analysis of loss-of-function alleles and a gain-of-function allele of atf-7, combined with expression analysis of PMK-1–regulated genes and biochemical characterization of the interaction between ATF-7 and PMK-1, suggest that ATF-7 functions as a repressor of PMK-1–regulated genes that undergoes a switch to an activator upon phosphorylation by PMK-1. Whereas loss-of-function mutations in atf-7 can restore basal expression of PMK-1–regulated genes observed in the pmk-1 null mutant, the induction of PMK-1–regulated genes by pathogenic Pseudomonas aeruginosa PA14 is abrogated. The switching modes of ATF-7 activity, from repressor to activator in response to activated PMK-1 p38 MAPK, are reminiscent of the mechanism of regulation mediated by the corresponding ancestral Sko1p and Hog1p proteins in the yeast response to osmotic stress. Our data point to the regulation of the ATF2/ATF7/CREB5 family of transcriptional regulators by p38 MAPK as an ancient conserved mechanism for the control of innate immunity in metazoans, and suggest that ATF2/ATF7 may function in a similar manner in the regulation of mammalian innate immunity. We have investigated mechanisms of how the soil nematode Caenorhabditis elegans interacts with pathogenic bacteria. Previously, we have established that a conserved PMK-1 p38 mitogen-activated protein kinase (MAPK) pathway regulates immunity in C. elegans, establishing the conservation of key innate immune signaling pathways of mammals in the immune response of C. elegans. Whereas multiple proteins have been identified as potential targets of p38 MAPK in immunity, the identification of physiological substrates of p38 MAPK in mammalian organisms has been challenging. Here, using a forward genetic approach to identify downstream regulators of the C. elegans innate immune response, we have characterized the transcription factor ATF-7, a conserved member of the basic-region leucine zipper (bZIP) transcription factor family orthologous to mammalian ATF2. We find that ATF-7 functions as a transcriptional regulator of PMK-1 MAPK–mediated innate immunity, functioning as a repressor of immune gene expression that undergoes a switch to an activator upon activation by PMK-1. Our data point to the regulation of the ATF2/ATF7/CREB5 family of transcriptional regulators by p38 MAPK as an ancient conserved mechanism for the control of innate immunity in metazoans and suggests a mechanism by which the protean effects of p38 MAPK on the mammalian innate immune response may be mediated.

From genetics to material to behavior: Introducing new genes into an organism can endow new biochemical functions or change the patterns of existing functions, but extending these manipulations to structure at the tissue level is challenging. Combining genetic engineering and polymer chemistry, Liu et al. directly leveraged complex cellular architectures of living organisms to synthesize, fabricate, and assemble bioelectronic materials (see the Perspective by Otto and Schmidt). An engineered enzyme expressed in genetically targeted neurons synthesized conductive polymers in tissues of freely moving animals. These polymers enabled modulation of membrane properties in specific neuron populations and manipulation of behavior in living animals. Science , this issue p. 1372 ; see also p. 1303

Endoplasmic reticulum (ER) stress activates the Unfolded Protein Response, a compensatory signaling response that is mediated by the IRE-1, PERK/PEK-1, and ATF-6 pathways in metazoans. Genetic studies have implicated roles for UPR signaling in animal development and disease, but the function of the UPR under physiological conditions, in the absence of chemical agents administered to induce ER stress, is not well understood. Here, we show that in Caenorhabditis elegans XBP-1 deficiency results in constitutive ER stress, reflected by increased basal levels of IRE-1 and PEK-1 activity under physiological conditions. We define a dynamic, temperature-dependent requirement for XBP-1 and PEK-1 activities that increases with immune activation and at elevated physiological temperatures in C. elegans. Our data suggest that the negative feedback loops involving the activation of IRE-1-XBP-1 and PEK-1 pathways serve essential roles, not only at the extremes of ER stress, but also in the maintenance of ER homeostasis under physiological conditions. Proteins destined for secretion outside of eukaryotic cells are trafficked through the endoplasmic reticulum (ER). Protein folding in the ER involves the activity of chaperones, as well as catalysis of post-translational modifications such as disulfide bond formation and glycosylation. When the folding capacity of the ER is exceeded, the resulting accumulation of misfolded proteins activates the Unfolded Protein Response (UPR), a conserved signaling response that functions to restore protein folding homeostasis in the ER. Genetic studies have established that the UPR is required for the development of specific cell types in mammals, such as antibody-secreting plasma cells, and recent studies implicate a critical role for UPR signaling in the pathogenesis of metabolic and inflammatory diseases. In this paper we show that innate immunity and elevated physiological temperatures each necessitate UPR activity for C. elegans survival. Furthermore, we show that, under physiological conditions of larval development, basal activity of the UPR is required for the maintenance of ER homeostasis. Our data support the idea not only that the UPR functions as a “stress response” pathway, protecting against the extremes of unfolded protein accumulation, but also that the UPR plays a more general role in animal physiology and development.

The translation initiation factor eIF3 is a multi-subunit protein complex that coordinates the assembly of the 43S pre-initiation complex in eukaryotes. Prior studies have demonstrated that not all subunits of eIF3 are essential for the initiation of translation, suggesting that some subunits may serve regulatory roles. Here, we show that loss-of-function mutations in the genes encoding the conserved eIF3k and eIF3l subunits of the translation initiation complex eIF3 result in a 40% extension in lifespan and enhanced resistance to endoplasmic reticulum (ER) stress in Caenorhabditis elegans. In contrast to previously described mutations in genes encoding translation initiation components that confer lifespan extension in C. elegans, loss-of-function mutations in eif-3.K or eif-3.L are viable, and mutants show normal rates of growth and development, and have wild-type levels of bulk protein synthesis. Lifespan extension resulting from EIF-3.K or EIF-3.L deficiency is suppressed by a mutation in the Forkhead family transcription factor DAF-16. Mutations in eif-3.K or eif-3.L also confer enhanced resistance to ER stress, independent of IRE-1-XBP-1, ATF-6, and PEK-1, and independent of DAF-16. Our data suggest a pivotal functional role for conserved eIF3k and eIF3l accessory subunits of eIF3 in the regulation of cellular and organismal responses to ER stress and aging. The translation initiation factor eIF3 is the largest and most complex of the eukaryotic initiation factors, comprising 13 subunits in metazoans, and this protein complex plays a critical role in scaffolding key interactions among multiple cofactors required for translation initiation. Both evolutionary and biochemical evidence demonstrate that the critical functionality of this complex can be mediated by a limited repertoire of eIF3 subunits, suggesting regulatory or modulatory roles for the remaining subunits. Here, we show that two conserved subunits of this complex, eIF3k and eIF3l, are nonessential for normal growth and viability in C. elegans, and that in their absence, rates of bulk protein translation are not diminished. Deletion of either eIF3k or eIF3l confers a dramatic extension in lifespan and also improves protein-folding homeostasis of the endoplasmic reticulum in C. elegans. These data suggest that eIF3k and eIF3l may function in the regulation of cellular and organism physiology.