Nf1 RasGAP Inhibition of LIMK2 Mediates a New Cross-Talk between Ras and Rho Pathways

Background

Ras GTPases mediate numerous biological processes through their ability to cycle between an inactive GDP-bound form and an active GTP-bound form. Guanine nucleotide exchange factors (GEFs) favor the formation of the active Ras-GTP, whereas GTPase activating proteins (GAPs) promote the formation of inactive Ras-GDP. Numerous studies have established complex signaling cross-talks between Ras GTPases and other members of the superfamily of small GTPases. GEFs were thought to play a major role in these cross-talks. However, recently GAPs were also shown to play crucial roles in these processes. Among RasGAPs, Nf1 is of special interest. Nf1 is responsible for the genetic disease Neurofibromatosis type I, and recent data strongly suggest that this RasGAP connects different signaling pathways.

Methodology/Principal Findings

In order to know if the RasGAP Nf1 might play a role in connecting Ras GTPases to other small GTPase pathways, we systematically looked for new partners of Nf1, by performing a yeast two-hybrid screening on its SecPH domain. LIMK2, a major kinase of the Rho/ROCK/LIMK2/cofilin pathway, was identified in this screening. We confirmed this interaction by co-immunoprecipitation experiments, and further characterized it. We also demonstrated its specificity: the close related homolog of LIMK2, LIMK1, does not interact with the SecPH domain of Nf1. We then showed that SecPH partially inhibits the kinase activity of LIMK2 on cofilin. Our results furthermore suggest a precise mechanism for this inhibition: in fact, SecPH would specifically prevent LIMK2 activation by ROCK, its upstream regulator.

Conclusions/Significance

Although previous data had already connected Nf1 to actin cytoskeleton dynamics, our study provides for the first time possible detailed molecular requirements of this involvement. Nf1/LIMK2 interaction and inhibition allows to directly connect neurofibromatosis type I to actin cytoskeleton remodeling, and provides evidence that the RasGAP Nf1 mediates a new cross-talk between Ras and Rho signaling pathways within the superfamily of small GTPases.     

Salmonella enterica Flagellin Is Recognized via FLS2 and Activates PAMP-Triggered Immunity in Arabidopsis thaliana

Infections with Salmonella enterica belong to the most prominent causes of food poisoning and infected fruits and vegetables represent important vectors for salmonellosis. Recent evidence indicates that plants recognize S. enterica and raise defense responses. Nonetheless, the molecular mechanisms controlling the interaction of S. enterica with plants are still largely unclear. Here, we show that flagellin from S. enterica represents a prominent pathogenassociated molecular pattern （PAMP） in Arabidopsis thaliana, which induces PAMP-triggered immunity （PTI） via the recognition of the fig22 domain by the receptor kinase FLS2. The Arabidopsis fls2 mutant shows reduced though not abolished PTI activation, indicating that plants rely also on recognition of other S. enterica PAMPs. Interestingly, the S. enterica type III secretion system （T3SS） mutant prgH- induced stronger defense gene expression than wild-type bacteria in Arabidopsis, suggesting that T3SS effectors are involved in defense suppression. Furthermore, we observe that S. enterica strains show variation in the fig22 epitope, which results in proteins with reduced PTI-inducing activity. Altogether, these results show that S. enterica activates PTI in Arabidopsis and suggest that, in order to accomplish plant colonization, S. enterica evolved strategies to avoid or suppress PTI.     

A comparison of the metabolic response to abdominal obesity in two Canadian Inuit and First Nations population

Inuit and Cree populations are known for high obesity rates despite markedly different rates of type 2 diabetes (T2DM). To document this apparent discrepancy we evaluated the impact of body size parameters and fasting insulin (FI) on several T2DM risk factors among Inuit and Cree populations (Québec, Canada). A total of 1,104 adults (≥18 years) Inuit and Cree individuals participated in a cross-sectional investigation. Interestingly, across both genders, across all levels of waist circumference (WC), Inuit showed lower levels of FI (age-adjusted, P < 0.0001) and fasting glucose (P < 0.0001) than Cree individuals. In both groups, a comparison of multi-sample structural equation models confirmed the predominant influence of WC on other traditional risk factors, compared to BMI. A preponderant direct impact of WC was observed on blood pressure (BP) parameters (P < 0.0001), high-density lipoprotein cholesterol (HDL-C) (P < 0.0001), and FI (P < 0.0001). Fasting glucose level appear to be influenced by WC via FI in both ethnic groups (P < 0.0001), while triacylglycerol (TAG) level was predominantly impacted by WC via FI, but only in Cree individuals (P < 0.0001). The main ethnic difference found was the strength of the impact of WC on FI, which was considerably higher among the Cree (λ = 2.4, P < 0.0001) than the Inuit (λ = 1.8, P < 0.0001). These results confirm the predominant role of abdominal adiposity in the complex and tenuous links of different traditional T2DM determinants. However, the ethnic difference in the impact of abdominal obesity on insulin levels across all WCs needs to be explored further.     

Seawater temperature, Gambierdiscus spp. variability and incidence of ciguatera poisoning in French Polynesia

In the context of global warming and climate change, ciguatera disease is put forward as an indicator of environmental disturbance. However, to validate this indicator, some unknown parameters such as the delay between environmental perturbation and outbreaks of ciguatera need to be investigated. The main goal of this study was to investigate the temporal link between the growth of Gambierdiscus spp., and one of its influencing factors and the declared cases of ciguatera disease in humans. Algal cell density and seawater temperature (SWT) were recorded monthly from February 1993 to December 2001 on the Atimaono barrier reef of Tahiti Island. Reports of ciguatera cases were obtained from three community health clinics near the study sites. The autoregressive integrated moving average model (ARIMA) shows: (1) SWT were positively associated with Gambierdiscus spp. growth at a lagtime of 13 and 17 months (p < 0.001); (2) Gambierdiscus spp. growth measured at a given time is related to a peak number of cases of ciguatera recorded 3 months after peak densities of this dinoflagellate (p < 0.001). These results allow the construction of a predictive model of the temporal link between ciguatera disease in humans and its etiologic agent: Gambierdiscus spp. This model constructed by using 1993–1999 data, then validated by 2000–2001 data, demonstrates an appreciable ability to predict changes in the incidence of ciguatera disease following algae blooms.     

Tissue-specific responses of lipoprotein lipase to dietary macronutrient composition as a predictor of weight gain over 4 years

This study evaluated if the effect of dietary macronutrient composition on adipose tissue lipoprotein lipase (ATLPL) and skeletal muscle lipoprotein lipase (SMLPL) predicted the long-term (over 4 years) changes in body weight and composition in free-living adults. Using a crossover design, 39 healthy subjects (n = 24 normal weight, n = 7 overweight, n = 8 obese) each followed a 2-week isocaloric high-carbohydrate (HC; 55% CHO:25% fat) and high-fat (HF; 30% CHO:50% fat) diet. On day 15 of each diet, biopsies were performed in the fasted state and 6 h after a meal. Body weight and composition were measured annually over 4 years. The outcomes for body weight, fat mass and % body fat were assessed using a linear two-stage mixed model. The mean (±SEM) increase in body weight and fat mass over 4 years was 0.29 ± 0.15 kg/year (P = 0.063) and 0.31 ± 0.15 kg/year (P = 0.051), respectively. The most consistent predictors of future body weight and fat changes were the ΔATLPL and ΔSMLPL responses (0-6 h) to a HC diet/meal. For the HC diet/meal, the subjects who had an increase in ATLPL activity/cell gained more % body fat over 4 years (P = 0.006) whereas subjects who had a decrease in SMLPL activity/g also had an increase in fat mass (P = 0.021). No significant relationships were observed between fasting ATLPL and SMLPL or enzyme responses to meals and any of the outcomes following the HF diet. In free-living adults the variability in tissue-specific lipoprotein lipase (LPL) responsiveness to a HC diet/meal predicts longitudinal changes in body composition.