Mitogen-activated protein kinases (MAPKs) are integral to the mechanisms by which cells respond to physiological stimuli and to a wide variety of environmental stresses. MAPK cascades can be inactivated at the MAPK activation step by members of the MAPK phosphatase (MKP) family. However, the components that act in MKP-regulated pathways have not been well characterized in the context of whole organisms. Here we characterize the Caenorhabditis elegans vhp-1 gene, encoding an MKP that acts preferentially on the c-Jun N-terminal kinase (JNK) and p38 MAPKs. We found that animals defective in vhp-1 are arrested during larval development. This vhp-1 defect is suppressed by loss-of-function mutations in the kgb-1, mek-1, and mlk-1 genes encoding a JNK-like MAPK, an MKK7-type MAPKK, and an MLK-type MAPKKK, respectively. The genetic and biochemical data presented here demonstrate a critical role for VHP-1 in the KGB-1 pathway. Loss-of-function mutations in each component in the KGB-1 pathway result in hypersensitivity to heavy metals. These results suggest that VHP-1 plays a pivotal role in the integration and fine-tuning of the stress response regulated by the KGB-1 MAPK pathway.

The p38 and JNK classes of mitogen-activated protein kinases (MAPKs) have evolutionarily conserved roles in the control of cellular responses to microbial and abiotic stresses. The mechanisms by which crosstalk between distinct p38 and c-Jun N-terminal kinase (JNK) MAPK pathways occurs with resultant integration of signaling information have been difficult to establish, particularly in the context of whole organism physiology. In Caenorhabditis elegans a PMK-1 p38 MAPK pathway is required for resistance to bacterial infection, and a KGB-1 JNK-like MAPK pathway has recently been shown to mediate resistance to heavy metal stress. Here, we show that two components of the KGB-1 pathway, MEK-1 MAPK kinase (MAPKK), a homolog of mammalian MKK7, and VHP-1 MAPK phosphatase (MKP), a homolog of mammalian MKP7, also regulate pathogen resistance through the modulation of PMK-1 activity. The regulation of p38 and JNK-like MAPK pathways mediating immunity and heavy metal stress by common MAPKK and MKP signaling components suggests pivotal roles for MEK-1 and VHP-1 in the integration of diverse stress signals contributing to pathogen resistance in C. elegans. In addition, these data point to mechanisms in multicellular organisms by which signals transduced by distinct MAPK pathways may be subject to physiological integration at the level of regulation of MAPK activity by MAPKKs and MKPs. M itogen-activated protein kinase (MAPK) signaling pathways serve as transducers of extracellular stimuli that allow cellular adaptation to changes in environment. MAPK pathways are highly evolutionarily conserved and play key roles in many diverse physiological processes, including development, growth and proliferation, stress responses, and immunity. MAPK activation generally involves the phosphorylation of Thr and Tyr residues in a signature T-X-Y activation domain motif by a dual-specificity MAPK kinase (MAPKK) (1). Whereas the genetic characterization of MAPK activation in yeast has shown a one-to-one correspondence between a particular MAPKK and a cognate MAPK (2), studies of MAPK activation in multicellular organisms suggest that at least two MAPKKs can function as activators of a MAPK (1), although the functional significance of the ability of multiple MAPKKs to activate a single MAPK remains unclear. Negative regulatory elements of MAPK signaling have been less well characterized. Many classes of phosphatases, including Ser͞Thr phosphatases, Tyr phosphatases, and dual-specificity MAPK phosphatases (MKPs), have been implicated in inhibition of MAPK signaling pathways (3).The MAPK signaling cassette represents perhaps the most ancient of evolutionarily conserved pathways of immunity, being conserved from plants to mammals (4, 5). Two classes of MAPKs, the c-Jun N-terminal kinase (JNK) and p38 MAPK, function as key mediators of stress and immune signaling in mammals (1, 5). The MKK4 and MKK7 MAPKKs have been shown to activate JNK (6-13), and the MKK3 and MKK6 MAPKKs serve as the major activators of p38 MAPK (14-21).Genetic analysis in mic...

The ability of neurons to undergo regenerative growth after injury is governed by cell-intrinsic and cell-extrinsic regeneration pathways. These pathways represent potential targets for therapies to enhance regeneration. However, the signaling pathways that orchestrate axon regeneration are not well understood. In Caenorhabditis elegans, the Jun N-terminal kinase (JNK) and p38 MAP kinase (MAPK) pathways are important for axon regeneration. We found that the C. elegans SVH-1 growth factor and its receptor, SVH-2 tyrosine kinase, regulate axon regeneration. Loss of SVH-1-SVH-2 signaling resulted in a substantial defect in the ability of neurons to regenerate, whereas its activation improved regeneration. Furthermore, SVH-1-SVH-2 signaling was initiated extrinsically by a pair of sensory neurons and functioned upstream of the JNK-MAPK pathway. Thus, SVH-1-SVH-2 signaling via activation of the MAPK pathway acts to coordinate neuron regeneration response after axon injury.

Escherichia coli contains a major cold shock protein, CspA (or CS7.4), whose production is predominantly induced at low temperatures. This bacterium is known to possess five additional genes, each encoding a protein highly similar to CspA (referred to as the CspA family). Here we identified a gene that encodes a cold-shockinducible analog of CspA and CspB. This newly cloned cspG gene is located at 22 min on the E. coli genetic map, apart from the other cspA family genes. Its gene product (70 amino acids) is 73 and 77% identical to CspA (70 amino acids) and CspB (71 amino acids), respectively. Analyses of a cspG-lacZ transcriptional fusion and Northern (RNA) hybridization revealed that cspG is a low-temperature-responsive gene. Its low-temperatureinducible promoters were determined, and the results indicated that the cspG sequence is highly similar to both the cspA and cspB sequences not only in the coding regions but also in the 5-upstream noncoding regions surrounding their own promoters.In Escherichia coli, temperature shifts from the normal range (20 to 37ЊC) to a temperature of above 40ЊC or below 20ЊC elicit pronounced physiological changes in growing cells. The best-characterized one is the so-called heat shock response (3). The heat shock proteins have become central to the study of the correct folding of nascent and/or unfolded polypeptides (1). Studies have also been initiated to elucidate the effects of a downshift in growth temperature on E. coli physiology. After the downshift, like heat shock proteins, a set of cold shock proteins (CSPs) were found to be produced at rates higher than those at the normal temperatures (7).

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Accumulation of unfolded proteins in the lumen of the endoplasmic reticulum (ER) causes ER stress. Snf1, the Saccharomyces cerevisiae ortholog of AMP–activated protein kinase (AMPK), plays a crucial role in the response to various environmental stresses. However, the role of Snf1 in ER stress response remains poorly understood. In this study, we characterize Snf1 as a negative regulator of Hog1 MAPK in ER stress response. The snf1 mutant cells showed the ER stress resistant phenotype. In contrast, Snf1-hyperactivated cells were sensitive to ER stress. Activated Hog1 levels were increased by snf1 mutation, although Snf1 hyperactivation interfered with Hog1 activation. Ssk1, a specific activator of MAPKKK functioning upstream of Hog1, was induced by ER stress, and its induction was inhibited in a manner dependent on Snf1 activity. Furthermore, we show that the SSK1 promoter is important not only for Snf1-modulated regulation of Ssk1 expression, but also for Ssk1 function in conferring ER stress tolerance. Our data suggest that Snf1 downregulates ER stress response signal mediated by Hog1 through negatively regulating expression of its specific activator Ssk1 at the transcriptional level. We also find that snf1 mutation upregulates the unfolded protein response (UPR) pathway, whereas Snf1 hyperactivation downregulates the UPR activity. Thus, Snf1 plays pleiotropic roles in ER stress response by negatively regulating the Hog1 MAPK pathway and the UPR pathway.

Hepatic butyltin concentrations were determined in 63 cetaceans belonging to 14 species and four pinnipeds belonging to two species collected from North Pacific and Asian coastal waters. Butyltin compounds (BTs) including tributyltin (TBT), dibutyltin (DBT), and monobutyltin (MBT) were detected in almost all the liver samples suggestive of its worldwide distribution. The elevated residues detected in coastal species and low concentrations found in off-shore species indicate a high degree of butyltin contamination in coastal waters than in the open sea. Mammals inhabiting waters of developed nations were found to contain higher BT concentrations compared with those collected from the waters proximal to developing countries. These observations strongly suggest serious BT contamination in the waters of developed countries than in developing nations at present. Among the samples collected off Japanese coastal waters, lower BT concentrations were found in pinnipeds compared with the cetaceans, suggestive of a possible difference in degradation capacities and excretory moulting between these two groups of animals. The estimated concentration ratio of BT in the liver of killer whale fetus to its pregnant mother was relatively low (0.015), indicative that transplacental transfer of BTs from the mother to her fetus is a deal less. Among the BT breakdown products, DBT was predominant in most of the liver samples analyzed, followed by TBT and MBT.

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