Thiocillin contributes to the ecological fitness of Bacillus cereus ATCC 14579 during interspecies interactions with Myxococcus xanthus.

The soil-dwelling delta-proteobacterium Myxococcus xanthus is a model organism to study predation and competition. M. xanthus preys on a broad range of bacteria mediated by lytic enzymes, exopolysaccharides, Type-IV pilus-based motility, and specialized metabolites. Competition between M. xanthus and prey bacterial strains with various specialized metabolite profiles indicates a range of fitness, suggesting that specialized metabolites contribute to prey survival. To expand our understanding of how specialized metabolites affect predator-prey dynamics, we assessed interspecies interactions between M. xanthus and two strains of Bacillus cereus. While strain ATCC 14579 resisted predation, strain T was found to be highly sensitive to M. xanthus predation. The interaction between B. cereus ATCC 14579 and M. xanthus appears to be competitive, resulting in population loss for both predator and prey. Genome analysis revealed that ATCC 14579 belongs to a clade that possesses the biosynthetic gene cluster for production of thiocillins, whereas B. cereus strain T lacks those genes. Further, purified thiocillin protects B. cereus strains unable to produce this specialized metabolite, strengthening the finding that thiocillin protects against predation and contributes to the ecological fitness of B. cereus ATCC 14579. Lastly, strains that produce thiocillin appear to confer some level of protection to their own antibiotic by encoding an additional copy of the L11 ribosomal protein, a known target for thiopeptides. This work highlights the importance of specialized metabolites affecting predator-prey dynamics in soil microenvironments.

Reading Fluency in Chinese Children With Reading Disabilities and/or ADHD: A Key Role for Morphology.

The Triangle Model of Reading proposes that phonology, orthography, and semantics are crucial to understand word reading and reading disability (RD). Morphology has been added as a binding agent to this model. However, it is unclear how these variables relate to word reading in children with attention deficit/hyperactivity disorder (ADHD) or comorbid ADHD and RD (ADHD+RD). This study examined the performance of Chinese children with RD, ADHD, or ADHD+RD in phonology, orthography, semantics, and morphology, and investigated whether morphology made an additional contribution beyond the other skills in explaining word reading fluency. Participants were 151 Grade 1 to 3 Chinese students: RD (n = 31), ADHD (n = 43), ADHD+RD (n = 27), and typically developing controls (TD, n = 50). Results indicated that children with ADHD+RD (a) showed similar performance to RD and ADHD in tone awareness, orthographic legality, and homophone morpheme awareness; (b) had similar performance to RD but worse than ADHD in phonology, semantics, and morpheme production; and (c) had more severe deficits than RD and ADHD in orthographic reversal, morpheme identification, and homograph awareness. Morphology significantly predicted word reading fluency beyond the other skills, and its predictive effect was more salient for ADHD+RD, ADHD, and TD. The findings provide evidence of both shared and additive effects of RD and ADHD. Morphology may be an important diagnostic factor in identifying Chinese reading and behavioral deficit groups and a worthwhile target for intervention.

The summary writing performance of bilingual learners with reading difficulties.

writing is an important tactic for learning from text and the summaries provide information on students' comprehension and learning processes. We investigated the nature of the summaries produced by bilingual adolescents, and whether their summaries were related to their reading abilities in their first and second languages. In each language, we examined the performance of students identified as typically developing, poor decoders, or poor comprehenders. Participants were 246 grade 8 students enrolled in English immersion programs in China. Measures included English word reading and reading comprehension, Chinese word reading and reading comprehension, and nonverbal ability. Students' text-absent summaries of an English 254-word expository passage were analyzed for the number of themes, main ideas, important details, and unimportant details. Using latent profile analysis, participants were identified as typical readers (TR, n = 123), poor decoders (PD, n = 74), or poor comprehenders (PC, n = 49) in English, and TR (n = 129), PD (n = 74), or PC (n = 43) in Chinese, based on word reading and reading comprehension in both English and Chinese. MANCOVA results showed that after controlling for nonverbal ability, in the English-defined groups, the TR group outperformed PD and PC on themes, main ideas, and important details; in the Chinese-defined groups, the TR group outperformed PD and PC on themes, TR performed better than PC on main ideas and important details, and PD outperformed PC on main ideas. Discussion focuses on the difficulties faced by bilingual students with reading difficulties and on the potential of summary writing instruction to improve their comprehension and learning processes.

A single, peri-operative antibiotic can persistently alter the post-operative gut microbiome after Roux-en-Y gastric bypass.

INTRODUCTION: Roux-en-Y gastric bypass (RYGB) significantly alters the gut microbiome and may be a mechanism for post-operative cardiovascular disease improvement. We have previously found an association between the class of peri-operative, intravenous antibiotic administered at the time of RYGB and the resolution rate of hypertension suggesting the gut microbiome as a mechanism. In this study, we performed a prospective study of RYGB to determine if a single intravenous antibiotic could alter the gastrointestinal microbial composition. METHODS: Patients undergoing RYGB were randomized to a single, peri-operative antibiotic of intravenous cefazolin (n = 8) or clindamycin (n = 8). Stool samples were collected from four-time points: 2 weeks pre-op (- 2w), 2 days pre-op (- 2d), 2 weeks post-op (+ 2w) and 3 months post-op (+ 3m). Stool samples were processed for genomic DNA followed by Illumina 16S rRNA gene sequencing and shotgun metagenomic sequencing (MGS). RESULTS: A total of 60 stool samples (- 2w, n = 16; - 2d, n = 15; + 2w, n = 16; + 3m, n = 13) from 16 patients were analyzed. 87.5% of patients were female with an average age of 48.6 ± 12.2 years and pre-operative BMI of 50.9 ± 23.3 kg/m2. RYGB induced statistically significant differences in alpha and beta diversity. There were statistically significant differences in alpha diversity at + 2w and beta diversity at + 3m due to antibiotic treatment. MGS revealed significantly distinct gut microbiota with 11 discriminatory metagenomic assembled genomes driven by antibiotic treatment at 3 months post-op, including increased Bifidobacterium spp. with clindamycin. CONCLUSION: RYGB induces significant changes in the gut microbiome at 2 weeks that are maintained 3 months after surgery. However, the single peri-operative dose of antibiotic administered at the time of RYGB induces unique and persisting changes to the gut microbiome that are antibiotic-specific. Increased Bifidobacterium spp. with clindamycin administration may improve the metabolic efficacy of RYGB when considering gut-microbiome driven mechanisms for blood pressure resolution.

Methods for the Comprehensive in vivo Analysis of Energy Flux, Fluid Homeostasis, Blood Pressure, and Ventilatory Function in Rodents.

Cardiovascular disease represents the leading cause of death in the United States, and metabolic diseases such as obesity represent the primary impediment to improving cardiovascular health. Rodent (mouse and rat) models are widely used to model cardiometabolic disease, and as a result, there is increasing interest in the development of accurate and precise methodologies with sufficiently high resolution to dissect mechanisms controlling cardiometabolic physiology in these small organisms. Further, there is great utility in the development of centralized core facilities furnished with high-throughput equipment configurations and staffed with professional content experts to guide investigators and ensure the rigor and reproducibility of experimental endeavors. Here, we outline the array of specialized equipment and approaches that are employed within the Comprehensive Rodent Metabolic Phenotyping Core (CRMPC) and our collaborating laboratories within the Departments of Physiology, Pediatrics, Microbiology & Immunology, and Biomedical Engineering at the Medical College of Wisconsin (MCW), for the detailed mechanistic dissection of cardiometabolic function in mice and rats. We highlight selected methods for the analysis of body composition and fluid compartmentalization, electrolyte accumulation and flux, energy accumulation and flux, physical activity, ingestive behaviors, ventilatory function, blood pressure, heart rate, autonomic function, and assessment and manipulation of the gut microbiota. Further, we include discussion of the advantages and disadvantages of these approaches for their use with rodent models, and considerations for experimental designs using these methods.

Nicotinamide Riboside-Conditioned Microbiota Deflects High-Fat Diet-Induced Weight Gain in Mice.

The gut microbiome plays an essential role in host energy homeostasis and influences the development of obesity and related conditions. Studies demonstrate that nicotinamide riboside (NR) supplementation for diet-induced obesity (DIO) reduces weight gain and increases energy expenditure in mice. NR is a vitamin B3 derivative and an NAD+ precursor with potential for treating human diseases arising from mitochondrial degeneration, including obesity and type 2 diabetes. Gut bacteria produce vitamin B3 in the colon and are capable of salvaging and metabolizing vitamin B3 and its derivatives. However, it is unknown how dietary supplementation of NR alters the microbiome and if those alterations contribute to deflection of weight gain. In this study, we fed C57BL/6J male mice a high-fat diet (HFD) supplemented with or without NR and performed a fecal material transfer (FMT) to establish a link between NR-conditioned microbiota and NR-induced deflection of weight gain. FMT from NR-treated donors to naive mice fed a HFD was sufficient to deflect weight gain by increasing energy expenditure. We also investigated the effects of NR on the microbiome by using metagenomics sequencing. We found that NR-treated mice displayed an altered gut microbial composition relative to controls and that fecal transplant resulted in a distinct functional metabolic profile characterized by enrichment of butyrate-producing Firmicutes. NR-treated donors and subsequent FMT recipients share a similar enrichment of metagenomic biomarkers relative to controls. These findings suggest that microbial factors contribute to the beneficial effects of dietary NR supplementation, which may be useful to enhance the therapeutic effects of NR. IMPORTANCE With obesity and type 2 diabetes (T2D) at epidemic levels, we need to understand the complex nature of these diseases to design better therapeutics. The underlying causes of both obesity and T2D are complex, but both are thought to develop, in part, based on contributions from the gut microbiota. Nicotinamide riboside is a gut-derived vitamin B3 derivative and NAD+ precursor which has the potential to treat and prevent metabolic disorders by ameliorating mitochondrial dysfunction. Understanding how NR affects the gut microbiome and whether NR-conditioned microbiota contributes to weight loss in the host would (i) improve diagnosis and treatments for obesity and other metabolic pathologies, (ii) tailor treatments to satisfy the needs of each individual moving toward the future of precision medicine, and (iii) benefit other scientific fields that currently investigate the effects of NR in other disease pathologies.

Profiles of Poor Decoders, Poor Comprehenders, and Typically Developing Readers in Adolescents Learning English as a Second Language.

This study examined (a) the identification of various reading groups across languages in Chinese (L1) adolescents learning English as a second language (ESL), in terms of their word-reading and reading comprehension skills, (b) overlap in reading group membership across languages, and (c) the performance of the various reading groups on reading-related language comprehension measures in English. The participants were 246 eighth-grade students from an English-immersion program in a middle school in China. Latent profile analysis identified three reading groups in each language: (a) a typically developing reader group with average or above-average word-reading and reading comprehension, (b) a group with poor decoding/word-reading skills and weak reading comprehension, and (c) a group with poor reading comprehension in the absence of poor decoding/word reading. The overlap in profile characteristics across languages for typically developing readers and poor decoders was high (about 68% for typically developing readers and 54% for poor decoders), whereas the overlap for being poor comprehenders in each language was moderate (about 37%). Furthermore, poor decoders in either language performed more poorly than the typically developing and poor comprehender groups on word reading in the other language, while poor comprehenders in either language performed more poorly than the typically developing and poor decoder groups on reading comprehension in the other language. The comparison of the reading groups' performance on English reading-related language comprehension measures showed that poor comprehenders and poor decoders performed worse than typically developing readers. Implications for identification and instruction of ESL children with reading difficulties are discussed.

Gut Microbiota Represent a Major Thermogenic Biomass.

Evidence supports various roles for microbial metabolites in the control of multiple aspects of host energy flux including feeding behaviors, digestive efficiency, and energy expenditure, but few studies have quantified the energy utilization of the biomass of the gut microbiota itself. Because gut microbiota exist in an anoxic environment, energy flux is expected to be anaerobic; unfortunately, commonly utilized O2/CO2 respirometry-based approaches are unable to detect anaerobic energy flux. To quantify the contribution of the gut microbial biomass to whole-animal energy flux, we examined the effect of surgical reduction of gut biomass in C57BL/6J mice via cecectomy and assessed energy expenditure using methods sensitive to anaerobic flux, including bomb and direct calorimetry. First, we determined that cecectomy caused an acceleration of weight gain over several months due to a reduction in combined total host plus microbial energy expenditure, as reflected by an increase in energy efficiency (ie, weight gained per calorie absorbed). Second, we determined that under general anesthesia, cecectomy caused immediate changes in heat dissipation that were significantly modified by short-term pretreatment with dietary or pharmaceutical interventions known to modify the microbiome, and confirmed that these effects were undetectable by respirometry. We conclude that while the cecum only contributes approximately 1% of body mass in the mouse, this organ contributes roughly 8% of total resting energy expenditure, that this contribution is predominantly anaerobic, and that the composition and abundance of the cecal microbial contents can significantly alter its contribution to energy flux.

Dietary influences on the Dahl SS rat gut microbiota and its effects on salt-sensitive hypertension and renal damage.

AIM: Our previous studies have demonstrated the importance of dietary factors in the determination of hypertension in Dahl salt-sensitive (SS) rats. Since the gut microbiota has been implicated in chronic diseases like hypertension, we hypothesized that dietary alterations shift the microbiota to mediate the development of salt-sensitive hypertension and renal disease. METHODS: This study utilized SS rats from the Medical College of Wisconsin (SS/MCW) maintained on a purified, casein-based diet (0.4% NaCl AIN-76A, Dyets) and from Charles River Laboratories (SS/CRL) fed a whole grain diet (0.75% NaCl 5L79, LabDiet). Faecal 16S rDNA sequencing was used to phenotype the gut microbiota. Directly examining the contribution of the gut microbiota, SS/CRL rats were administered faecal microbiota transfer (FMT) experiments with either SS/MCW stool or vehicle (Vehl) in conjunction with the HS AIN-76A diet. RESULTS: SS/MCW rats exhibit renal damage and inflammation when fed high salt (HS, 4.0% NaCl AIN-76A), which is significantly attenuated in SS/CRL. Gut microbiota phenotyping revealed distinct profiles that correlate with disease severity. SS/MCW FMT worsened the SS/CRL response to HS, evidenced by increased albuminuria (67.4 ± 6.9 vs 113.7 ± 25.0 mg/day, Vehl vs FMT, P = .007), systolic arterial pressure (158.6 ± 5.8 vs 177.8 ± 8.9 mmHg, Vehl vs FMT, P = .09) and renal T-cell infiltration (1.9-fold). Amplicon sequence variant (ASV)-based analysis of faecal 16S rDNA sequencing data revealed taxa that significantly shifted with FMT: Erysipelotrichaceae_2, Parabacteroides gordonii, Streptococcus alactolyticus, Bacteroidales_1, Desulfovibrionaceae_2, Ruminococcus albus. CONCLUSIONS: These data demonstrate that dietary modulation of the gut microbiota directly contributes to the development of Dahl SS hypertension and renal injury.

Developmental Trends of Visual Processing of Letters and Objects Using Naming Speed Tasks.

Studying the typical development of reading is key to understanding the precise deficits that underlie reading disabilities. An important correlate of efficient reading is the speed of naming arrays of simple stimuli such as letters and pictures. In this cross-sectional study, we examined developmental changes in visual processing that occurs during letter and object naming from childhood to early adulthood in terms of behavioral task efficiency, associated articulation and eye movement parameters, and the coordination between them, as measured by eye-voice span in both the spatial and temporal domains. We used naming speed (NS) tasks, in which participants were required to name sets of letters or simple objects as quickly and as accurately as possible. Single stimulus manipulations were made to these tasks to make the stimuli either more visually and/or phonologically similar to one another in order to examine how these manipulations affected task performance and the coordination between speech and eye movements. Across development there was an increased efficiency in speech and eye movement performance and their coordination in both the spatial and temporal domains. Furthermore, manipulations to the phonological and visual similarity of specific letter and object stimuli revealed that orthographic processing played a greater role than phonological processing in performance, with the contribution of phonological processing diminishing across development. This comprehensive typical developmental trajectory provides a benchmark for clinical populations to elucidate the nature of the cognitive dysfunction underlying reading difficulties.

Understanding the biological basis of dyslexia at a neural systems level.

We examined the naming speed performance of 18 typically achieving and 16 dyslexic adults while simultaneously recording eye movements, articulations and fMRI data. Naming speed tasks, which require participants to name a list of letters or objects, have been proposed as a proxy for reading and are thought to recruit similar reading networks in the left hemisphere of the brain as more complex reading tasks. We employed letter and object naming speed tasks, with task manipulations to make the stimuli more or less phonologically and/or visually similar. Compared to typically achieving readers, readers with dyslexia had a poorer behavioural naming speed task performance, longer fixation durations, more regressions and increased activation in areas of the reading network in the left-hemisphere. Whereas increased network activation was positively associated with performance in dyslexics, it was negatively related to performance in typically achieving readers. Readers with dyslexia had greater bilateral activation and recruited additional regions involved with memory, namely the amygdala and hippocampus; in contrast, the typically achieving readers additionally activated the dorsolateral prefrontal cortex. Areas within the reading network were differentially activated by stimulus manipulations to the naming speed tasks. There was less efficient naming speed behavioural performance, longer fixation durations, more regressions and increased neural activity when letter stimuli were both phonologically and visually similar. Discussion focuses on the differences in activation within the reading network, how they are related to behavioural task differences, and how progress in furthering the understanding of the relationship between behavioural performance and brain activity can change the overall trajectories of children with reading difficulties by contributing to both early identification and remediation processes.

Correction to: Predicting Arabic word reading: A cross-classified generalized random-effects analysis showing the critical role of morphology.

The original version of this article unfortunately contained the following errors.

The SrrAB two-component system regulates Staphylococcus aureus pathogenicity through redox sensitive cysteines.

Staphylococcus aureus infections can lead to diseases that range from localized skin abscess to life-threatening toxic shock syndrome. The SrrAB two-component system (TCS) is a global regulator of S. aureus virulence and critical for survival under environmental conditions such as hypoxic, oxidative, and nitrosative stress found at sites of infection. Despite the critical role of SrrAB in S. aureus pathogenicity, the mechanism by which the SrrAB TCS senses and responds to these environmental signals remains unknown. Bioinformatics analysis showed that the SrrB histidine kinase contains several domains, including an extracellular Cache domain and a cytoplasmic HAMP-PAS-DHp-CA region. Here, we show that the PAS domain regulates both kinase and phosphatase enzyme activity of SrrB and present the structure of the DHp-CA catalytic core. Importantly, this structure shows a unique intramolecular cysteine disulfide bond in the ATP-binding domain that significantly affects autophosphorylation kinetics. In vitro data show that the redox state of the disulfide bond affects S. aureus biofilm formation and toxic shock syndrome toxin-1 production. Moreover, with the use of the rabbit infective endocarditis model, we demonstrate that the disulfide bond is a critical regulatory element of SrrB function during S. aureus infection. Our data support a model whereby the disulfide bond and PAS domain of SrrB sense and respond to the cellular redox environment to regulate S. aureus survival and pathogenesis.

Achromobacter xylosoxidans Cellular Pathology Is Correlated with Activation of a Type III Secretion System.

Achromobacter xylosoxidans is increasingly recognized as a colonizer of cystic fibrosis (CF) patients, but the role that A. xylosoxidans plays in pathology remains unknown. This knowledge gap is largely due to the lack of model systems available to study the toxic potential of this bacterium. Recently, a phospholipase A2 (PLA2) encoded by a majority of A. xylosoxidans genomes, termed AxoU, was identified. Here, we show that AxoU is a type III secretion system (T3SS) substrate that induces cytotoxicity to mammalian cells. A tissue culture model was developed showing that a subset of A. xylosoxidans isolates from CF patients induce cytotoxicity in macrophages, suggestive of a pathogenic or inflammatory role in the CF lung. In a toxic strain, cytotoxicity is correlated with transcriptional activation of axoU and T3SS genes, demonstrating that this model can be used as a tool to identify and track expression of virulence determinants produced by this poorly understood bacterium.

Predicting Arabic word reading: A cross-classified generalized random-effects analysis showing the critical role of morphology.

The distinctive features of the Arabic language and orthography offer opportunities to investigate multiple word characteristics at the item level. The aim of this paper was to model differences in word reading at the item level among 3rd grade native Arabic-speaking children (n = 303) using cross-classified generalized random-effects (CCGRE) analysis. The participants read 80 vowelized words that varied in multiple elements that may contribute to their decodability: number of letters, number of syllables, number of morphemes, ligaturing (connectivity), semantics (concrete vs. abstract), orthographic frequency, root type frequency, and part of speech. Morphological awareness (MA) was included as a person-level predictor. Results of individual models showed that MA, number of letters, number of syllables, number of morphemes, number of ligatures, orthographic frequency, and part of speech were significantly related to the probability of a correct response. However, when all predictors were entered simultaneously, only MA and number of morphemes remained significant. These results underscore the important role of morphology in the lexical structure of Arabic words and in Arabic word reading. Discussion focuses on the role of morphology in Arabic reading and the implications for intervention to improve word recognition in children learning to read Arabic.

Examining the neural and cognitive processes that underlie reading through naming speed tasks.

We combined fMRI with eye tracking and speech recording to examine the neural and cognitive mechanisms that underlie reading. To simplify the study of the complex processes involved during reading, we used naming speed (NS) tasks (also known as rapid automatized naming or RAN) as a focus for this study, in which average reading right-handed adults named sets of stimuli (letters or objects) as quickly and accurately as possible. Due to the possibility of spoken output during fMRI studies creating motion artifacts, we employed both an overt session and a covert session. When comparing the two sessions, there were no significant differences in behavioral performance, sensorimotor activation (except for regions involved in the motor aspects of speech production) or activation in regions within the left-hemisphere-dominant neural reading network. This established that differences found between the tasks within the reading network were not attributed to speech production motion artifacts or sensorimotor processes. Both behavioral and neuroimaging measures showed that letter naming was a more automatic and efficient task than object naming. Furthermore, specific manipulations to the NS tasks to make the stimuli more visually and/or phonologically similar differentially activated the reading network in the left hemisphere associated with phonological, orthographic and orthographic-to-phonological processing, but not articulatory/motor processing related to speech production. These findings further our understanding of the underlying neural processes that support reading by examining how activation within the reading network differs with both task performance and task characteristics.

Reading in Arabic: How Well Does the Standard Model Apply?

Purpose We investigated the cognitive and linguistic processes that underlie reading in Arabic in relation to a well-defined theoretical framework of reading and the factors that underlie reading. Method The sample was 201 (101 boys, 100 girls) 3rd-grade Arabic-speaking children. Children were administered measures of Vocabulary, Phonological Awareness (PA), Naming Speed, Orthographic Processing, Morphological Awareness (MA), Memory, Nonverbal Ability, and 5 reading outcomes. Hierarchical regression analyses were conducted for each of the 5 reading outcomes to investigate the predictors of children's reading. Results Each of the constructs explained unique variance when added to the model. In the final models, PA was the strongest predictor of all outcomes, followed by MA. In a follow-up analysis, participants were divided into good and poor decoders, based on their Pseudoword Reading scores. Good decoders outscored poor decoders on every measure. Within-group regression analyses indicated that poor decoders relied on more component processes than good decoders, suggesting a lack of automaticity. Variance in reading outcomes was better predicted for poor decoders than for good decoders. Conclusion These results indicate that standard predictors apply well to Arabic, showing the particular importance of PA and MA. Longitudinal and instructional studies are required to determine developmental patterns and ways to improve reading performance.

Suppressor mutations reveal an NtrC-like response regulator, NmpR, for modulation of Type-IV Pili-dependent motility in Myxococcus xanthus.

Two-component signaling systems (TCS) regulate bacterial responses to environmental signals through the process of protein phosphorylation. Specifically, sensor histidine kinases (SK) recognize signals and propagate the response via phosphorylation of a cognate response regulator (RR) that functions to initiate transcription of specific genes. Signaling within a single TCS is remarkably specific and cross-talk between TCS is limited. However, regulation of the flow of information through complex signaling networks that include closely related TCS remains largely unknown. Additionally, many bacteria utilize multi-component signaling networks which provide additional genetic and biochemical interactions that must be regulated for signaling fidelity, input and output specificity, and phosphorylation kinetics. Here we describe the characterization of an NtrC-like RR that participates in regulation of Type-IV pilus-dependent motility of Myxococcus xanthus and is thus named NmpR, NtrC Modulator of Pili Regulator. A complex multi-component signaling system including NmpR was revealed by suppressor mutations that restored motility to cells lacking PilR, an evolutionarily conserved RR required for expression of pilA encoding the major Type-IV pilus monomer found in many bacterial species. The system contains at least four signaling proteins: a SK with a protoglobin sensor domain (NmpU), a hybrid SK (NmpS), a phospho-sink protein (NmpT), and an NtrC-like RR (NmpR). We demonstrate that ΔpilR bypass suppressor mutations affect regulation of the NmpRSTU multi-component system, such that NmpR activation is capable of restoring expression of pilA in the absence of PilR. Our findings indicate that pilus gene expression in M. xanthus is regulated by an extended network of TCS which interact to refine control of pilus function.