The Eukaryotic Proteome Is Shaped by E3 Ubiquitin Ligases Targeting C-Terminal Degrons.

Degrons are minimal elements that mediate the interaction of proteins with degradation machineries to promote proteolysis. Despite their central role in proteostasis, the number of known degrons remains small, and a facile technology to characterize them is lacking. Using a strategy combining global protein stability (GPS) profiling with a synthetic human peptidome, we identify thousands of peptides containing degron activity. Employing CRISPR screening, we establish that the stability of many proteins is regulated through degrons located at their C terminus. We characterize eight Cullin-RING E3 ubiquitin ligase (CRL) complex adaptors that regulate C-terminal degrons, including six CRL2 and two CRL4 complexes, and computationally implicate multiple non-CRLs in end recognition. Proteome analysis revealed that the C termini of eukaryotic proteins are depleted for C-terminal degrons, suggesting an E3-ligase-dependent modulation of proteome composition. Thus, we propose that a series of "C-end rules" operate to govern protein stability and shape the eukaryotic proteome.

Ubiquitin-independent proteasomal degradation driven by C-degron pathways.

Although most eukaryotic proteins are targeted for proteasomal degradation by ubiquitination, a subset have been demonstrated to undergo ubiquitin-independent proteasomal degradation (UbInPD). However, little is known about the molecular mechanisms driving UbInPD and the degrons involved. Utilizing the GPS-peptidome approach, a systematic method for degron discovery, we found thousands of sequences that promote UbInPD; thus, UbInPD is more prevalent than currently appreciated. Furthermore, mutagenesis experiments revealed specific C-terminal degrons required for UbInPD. Stability profiling of a genome-wide collection of human open reading frames identified 69 full-length proteins subject to UbInPD. These included REC8 and CDCA4, proteins which control proliferation and survival, as well as mislocalized secretory proteins, suggesting that UbInPD performs both regulatory and protein quality control functions. In the context of full-length proteins, C termini also play a role in promoting UbInPD. Finally, we found that Ubiquilin family proteins mediate the proteasomal targeting of a subset of UbInPD substrates.

Elucidation of E3 ubiquitin ligase specificity through proteome-wide internal degron mapping.

The ubiquitin-proteasome system plays a critical role in biology by regulating protein degradation. Despite their importance, precise recognition specificity is known for a few of the 600 E3s. Here, we establish a two-pronged strategy for identifying and mapping critical residues of internal degrons on a proteome-scale in HEK-293T cells. We employ global protein stability profiling combined with machine learning to identify 15,800 peptides likely to contain sequence-dependent degrons. We combine this with scanning mutagenesis to define critical residues for over 5,000 predicted degrons. Focusing on Cullin-RING ligase degrons, we generated mutational fingerprints for 219 degrons and developed DegronID, a computational algorithm enabling the clustering of degron peptides with similar motifs. CRISPR analysis enabled the discovery of E3-degron pairs, of which we uncovered 16 pairs that revealed extensive degron variability and structural determinants. We provide the visualization of these data on the public DegronID data browser as a resource for future exploration.

The corepressors GPS2 and SMRT control enhancer and silencer remodeling via eRNA transcription during inflammatory activation of macrophages.

While the role of transcription factors and coactivators in controlling enhancer activity and chromatin structure linked to gene expression is well established, the involvement of corepressors is not. Using inflammatory macrophage activation as a model, we investigate here a corepressor complex containing GPS2 and SMRT both genome-wide and at the Ccl2 locus, encoding the chemokine CCL2 (MCP-1). We report that corepressors co-occupy candidate enhancers along with the coactivators CBP (H3K27 acetylase) and MED1 (mediator) but act antagonistically by repressing eRNA transcription-coupled H3K27 acetylation. Genome editing, transcriptional interference, and cistrome analysis reveals that apparently related enhancer and silencer elements control Ccl2 transcription in opposite ways. 4C-seq indicates that corepressor depletion or inflammatory signaling functions mechanistically similarly to trigger enhancer activation. In ob/ob mice, adipose tissue macrophage-selective depletion of the Ccl2 enhancer-transcribed eRNA reduces metaflammation. Thus, the identified corepressor-eRNA-chemokine pathway operates in vivo and suggests therapeutic opportunities by targeting eRNAs in immuno-metabolic diseases.

Mitochondrial Retrograde Signaling in Mammals Is Mediated by the Transcriptional Cofactor GPS2 via Direct Mitochondria-to-Nucleus Translocation.

As most of the mitochondrial proteome is encoded in the nucleus, mitochondrial functions critically depend on nuclear gene expression and bidirectional mito-nuclear communication. However, mitochondria-to-nucleus communication pathways in mammals are incompletely understood. Here, we identify G-Protein Pathway Suppressor 2 (GPS2) as a mediator of mitochondrial retrograde signaling and a transcriptional activator of nuclear-encoded mitochondrial genes. GPS2-regulated translocation from mitochondria to nucleus is essential for the transcriptional activation of a nuclear stress response to mitochondrial depolarization and for supporting basal mitochondrial biogenesis in differentiating adipocytes and brown adipose tissue (BAT) from mice. In the nucleus, GPS2 recruitment to target gene promoters regulates histone H3K9 demethylation and RNA POL2 activation through inhibition of Ubc13-mediated ubiquitination. These findings, together, reveal an additional layer of regulation of mitochondrial gene transcription, uncover a direct mitochondria-nuclear communication pathway, and indicate that GPS2 retrograde signaling is a key component of the mitochondrial stress response in mammals.

SkyPole-A method for locating the north celestial pole from skylight polarization patterns.

True north can be determined on Earth by three means: magnetic compasses, stars, and via the global navigation satellite systems (GNSS), each of which has its own drawbacks. GNSS are sensitive to jamming and spoofing, magnetic compasses are vulnerable to magnetic interferences, and the stars can be used only at night with a clear sky. As an alternative to these methods, nature-inspired navigational cues are of particular interest. Celestial polarization, which is used by insects such as Cataglyphis ants, can provide useful directional cues. Migrating birds calibrate their magnetic compasses by observing the celestial rotation at night. By combining these cues, we have developed a bioinspired optical method for finding the celestial pole during the daytime. This method, which we have named SkyPole, is based on the rotation of the skylight polarization pattern. A polarimetric camera was used to measure the degree of skylight polarization rotating with the Sun. Image difference processes were then applied to the time-varying measurements in order to determine the north celestial pole's position and thus the observer's latitude and bearing with respect to the true north.

GPS2 ameliorates cigarette smoking-induced pulmonary vascular remodeling by modulating the ras-Raf-ERK axis.

BACKGROUND: Mitogen-activated protein kinase (MAPK)signaling-mediated smoking-associated pulmonary vascular remodeling (PVR) plays an important role in the pathogenesis of group 3 pulmonary hypertension (PH). And G protein pathway suppressor 2 (GPS2) could suppress G-protein signaling such as Ras and MAPK, but its role in cigarette smoking -induced PVR (CS-PVR) is unclear. METHODS: An in vivo model of smoke-exposed rats was constructed to assess the role of GPS2 in smoking-induced PH and PVR. In vitro, the effects of GPS2 overexpression and silencing on the function of human pulmonary arterial smooth cells (HPASMCs) and the underlying mechanisms were explored. RESULTS: GPS2 expression was downregulated in rat pulmonary arteries (PAs) and HPASMCs after CS exposure. More importantly, CS-exposed rats with GPS2 overexpression had lower right ventricular systolic pressure (RVSP), right ventricular hypertrophy index (RVHI), and wall thickness (WT%) than those without. And enhanced proliferation and migration of HPASMCs induced by cigarette smoking extract (CSE) can be evidently inhibited by overexpressed GPS2. Besides, GPS2siRNA significantly enhanced the proliferation, and migration of HPASMCs as well as activated Ras and Raf/ERK signaling, while these effects were inhibited by zoledronic acid (ZOL). In addition, GPS2 promoter methylation level in rat PAs and HPASMCs was increased after CS exposure, and 5-aza-2-deoxycytidine (5-aza) inhibited CSE-induced GPS2 hypermethylation and downregulation in vitro. CONCLUSIONS: GPS2 overexpression could improve the CS-PVR, suggesting that GPS2 might serve as a novel therapeutic target for PH-COPD in the future.

Key Grazing Behaviours of Beef Cattle Identify Specific Genotypes of the Glutamate Metabotropic Receptor 5 Gene (GRM5).

Genotype-phenotype associations between the bovine genome and grazing behaviours measured over time and across contexts have been reported in the past decade, with these suggesting the potential for genetic control over grazing personalities in beef cattle. From the large array of metrics used to describe grazing personality behaviours (GP-behaviours), it is still unclear which ones are linked to specific genes. Our prior observational study has reported associations and trends towards associations between genotypes of the glutamate metabotropic receptor 5 gene (GRM5) and four GP-behaviours, yet the unbalanced representation of GRM5 genotypes occurring in observational studies may have limited the ability to detect associations. Here, we applied a subsampling technique to create a genotypically-balanced dataset in a quasi-manipulative experiment with free ranging cows grazing in steep and rugged terrain of New Zealand's South Island. Using quadratic discriminant analysis, two combinations of eleven GP-behaviours (and a total of fifteen behaviours) were selected to build an exploration model and an elevation model, respectively. Both models achieved ∼ 86% accuracy in correctly discriminating cows' GRM5 genotypes with the training dataset, and the exploration model achieved 85% correct genotype prediction of cows from a testing dataset. Our study suggests a potential pleiotropic effect, with GRM5 controlling multiple grazing behaviours, and with implications for the grazing of steep and rugged grasslands. The study highlights the importance of grazing behavioural genetics in cattle and the potential use of GRM5 markers to select individuals with desired grazing personalities and built herds that collectively utilize steep and rugged rangelands sustainably.

Comparing malaria risk exposure in rural Cambodia population using GPS tracking and questionnaires.

BACKGROUND: The Great Mekong Subregion has attained a major decline in malaria cases and fatalities over the last years, but residual transmission hotspots remain, supposedly fueled by forest workers and migrant populations. This study aimed to: (i) characterize the fine-scale mobility of forest-goers and understand links between their daily movement patterns and malaria transmission, using parasites detection via real time polymerase chain reaction (RT PCR) and the individual exposure to Anopheles bites by quantification of anti-Anopheles saliva antibodies via enzyme-linked immunosorbent assay; (ii) assess the concordance of questionnaires and Global Positioning System (GPS) data loggers for measuring mobility. METHODS: Two 28 day follow-ups during dry and rainy seasons, including a GPS tracking, questionnaires and health examinations, were performed on male forest goers representing the population at highest risk of infection. Their time spent in different land use categories and demographic data were analyzed in order to understand the risk factors driving malaria in the study area. RESULTS: Malaria risk varied with village forest cover and at a resolution of only a few kilometers: participants from villages outside the forest had the highest malaria prevalence compared to participants from forest fringe's villages. The time spent in a specific environment did not modulate the risk of malaria, in particular the time spent in forest was not associated with a higher probability to detect malaria among forest-goers. The levels of antibody response to Anopheles salivary peptide among participants were significantly higher during the rainy season, in accordance with Anopheles mosquito density variation, but was not affected by sociodemographic and mobility factors. The agreement between GPS and self-reported data was only 61.9% in reporting each kind of visited environment. CONCLUSIONS: In a context of residual malaria transmission which was mainly depicted by P. vivax asymptomatic infections, the implementation of questionnaires, GPS data-loggers and quantification of anti-saliva Anopheles antibodies on the high-risk group were not powerful enough to detect malaria risk factors associated with different mobility behaviours or time spent in various environments. The joint implementation of GPS trackers and questionnaires allowed to highlight the limitations of both methodologies and the benefits of using them together. New detection and follow-up strategies are still called for.

Human access constrains optimal foraging and habitat availability in an avian generalist.

Animals balance costs of antipredator behaviors with resource acquisition to minimize hunting and other mortality risks and maximize their physiological condition. This inherent trade-off between forage abundance, its quality, and mortality risk is intensified in human-dominated landscapes because fragmentation, habitat loss, and degradation of natural vegetation communities is often coupled with artificially enhanced vegetation (i.e., food plots), creating high-risk, high-reward resource selection decisions. Our goal was to evaluate autumn-winter resource selection trade-offs for an intensively hunted avian generalist. We hypothesized human access was a reliable cue for hunting predation risk. Therefore, we predicted resource selection patterns would be spatiotemporally dependent upon levels of access and associated perceived risk. Specifically, we evaluated resource selection of local-scale flights between diel periods for 426 mallards (Anas platyrhynchos) relative to wetland type, forage quality, and differing levels of human access across hunting and nonhunting seasons. Mallards selected areas that prohibited human access and generally avoided areas that allowed access diurnally, especially during the hunting season. Mallards compensated by selecting for high-energy and greater quality foraging patches on allowable human access areas nocturnally when they were devoid of hunters. Postseason selection across human access gradients did not return to prehunting levels immediately, perhaps suggesting a delayed response to reacclimate to nonhunted activities and thus agreeing with the assessment mismatch hypothesis. Last, wetland availability and human access constrained selection for optimal natural forage quality (i.e., seed biomass and forage productivity) diurnally during preseason and hunting season, respectively; however, mallards were freed from these constraints nocturnally during hunting season and postseason periods. Our results suggest risk-avoidance of human accessible (i.e., hunted) areas is a primary driver of resource selection behaviors by mallards and could be a local to landscape-level process influencing distributions, instead of forage abundance and quality, which has long-been assumed by waterfowl conservation planners in North America. Broadly, even an avian generalist, well adapted to anthropogenic landscapes, avoids areas where hunting and human access are allowed. Future conservation planning and implementation must consider management for recreational access (i.e., people) equally important as foraging habitat management for wintering waterfowl.

ExMove: An open-source toolkit for processing and exploring animal-tracking data in R.

Ongoing technological advances have led to a rapid increase in the number, type and scope of animal-tracking studies. In response, many software tools have been developed to analyse animal movement data. These tools generally focus on movement modelling, but the steps required to clean raw data files from different tracking devices have been largely ignored. Such pre-processing steps are often time-consuming and involve a steep learning curve but are crucial for the creation of high-quality, standardised and shareable data. Moreover, decisions made at this early stage can substantially influence subsequent analyses, and in the current age of reproducibility crisis, the transparency of this process is vital. Here we present an open-access, reproducible toolkit written in the programming language R for processing raw data files into a single cleaned data set for analyses and upload to online tracking databases (found here: https://github.com/ExMove/ExMove). The toolkit comprises well-documented and flexible code to facilitate data processing and user understanding, both of which can increase user confidence and improve the uptake of sharing open and reproducible code. Additionally, we provide an overview website (found here: https://exmove.github.io/) and a Shiny app to help users visualise tracking data and assist with parameter determination during data cleaning. The toolkit is generalisable to different data formats and device types, uses modern 'tidy coding' practices, and relies on a few well-maintained packages. Among these, we perform spatial manipulations using the package sf. Overall, by collating all required steps from data collection to archiving on open access databases into a single, robust pipeline, our toolkit provides a valuable resource for anyone conducting animal movement analyses and represents an important step towards increased standardisation and reproducibility in animal movement ecology.

Testing the information centre hypothesis in a multilevel society.

In various animal species conspecifics aggregate at sleeping sites. Such aggregations can act as information centres where individuals acquire up-to-date knowledge about their environment. In some species, communal sleeping sites comprise individuals from multiple groups, where each group maintains stable membership over time. We used GPS tracking to simultaneously record group movement in a population of wild vulturine guineafowl (Acryllium vulturinum) to investigate whether communal sleeping sites can facilitate the transfer of information among individuals across distinct groups. These birds live in large and stable groups that move both together and apart, often forming communal roosts containing up to five groups. We first test whether roosts provide the opportunity for individuals to acquire information from members of other groups by examining the spatial organization at roosts. The GPS data reveal that groups intermix, thereby providing an opportunity for individuals to acquire out-group information. We next conduct a field experiment to test whether naïve groups can locate novel food patches when co-roosting with knowledgeable groups. We find that co-roosting substantially increases the chances for the members of a naïve group to discover a patch known to individuals from other groups at the shared roost. Further, we find that the discovery of food patches by naïve groups subsequently shapes their space use and inter-group associations. We also draw on our long-term tracking to provide examples that demonstrate natural cases where communal roosting has preceded large-scale multi-group collective movements that extend into areas beyond the groups' normal ranges. Our findings support the extension of the information centre hypothesis to communal sleeping sites that consist of distinct social groups.

Validity of Mobility-Based Exposure Assessment of Air Pollution: A Comparative Analysis with Home-Based Exposure Assessment.

Air pollution exposure is typically assessed at the front door where people live in large-scale epidemiological studies, overlooking individuals' daily mobility out-of-home. However, there is limited evidence that incorporating mobility data into personal air pollution assessment improves exposure assessment compared to home-based assessments. This study aimed to compare the agreement between mobility-based and home-based assessments with personal exposure measurements. We measured repeatedly particulate matter (PM2.5) and black carbon (BC) using a sample of 41 older adults in the Netherlands. In total, 104 valid 24 h average personal measurements were collected. Home-based exposures were estimated by combining participants' home locations and temporal-adjusted air pollution maps. Mobility-based estimates of air pollution were computed based on smartphone-based tracking data, temporal-adjusted air pollution maps, indoor-outdoor penetration, and travel mode adjustment. Intraclass correlation coefficients (ICC) revealed that mobility-based estimates significantly improved agreement with personal measurements compared to home-based assessments. For PM2.5, agreement increased by 64% (ICC: 0.39-0.64), and for BC, it increased by 21% (ICC: 0.43-0.52). Our findings suggest that adjusting for indoor-outdoor pollutant ratios in mobility-based assessments can provide more valid estimates of air pollution than the commonly used home-based assessments, with no added value observed from travel mode adjustments.

Investigating young children's physical activity through time and place.

BACKGROUND: Previous research indicates the start of primary school (4-5-year-old) as an essential period for the development of children's physical activity (PA) patterns, as from this point, the age-related decline of PA is most often observed. During this period, young children are exposed to a wider variety of environmental- and social contexts and therefore their PA is influenced by more diverse factors. However, in order to understand children's daily PA patterns and identify relevant opportunities for PA promotion, it is important to further unravel in which (social) contexts throughout the day, PA of young children takes place. METHODS: We included a cross-national sample of 21 primary schools from the Startvaardig study. In total, 248 children provided valid accelerometer and global positioning (GPS) data. Geospatial analyses were conducted to quantify PA in (social) environments based on their school and home. Transport-related PA was evaluated using GPS speed-algorithms. PA was analysed at different environments, time-periods and for week- and weekend days separately. RESULTS: Children accumulated an average of 60 min of moderate-to-vigorous PA (MVPA), both during week- and weekend days. Schools contributed to approximately half of daily MVPA during weekdays. During weekends, environments within 100 m from home were important, as well as locations outside the home-school neighbourhood. Pedestrian trips contributed to almost half of the daily MVPA. CONCLUSIONS: We identified several social contexts relevant for children's daily MVPA. Schools have the potential to significantly contribute to young children's PA patterns and are therefore encouraged to systematically evaluate and implement parts of the school-system that stimulate PA and potentially also learning processes. Pedestrian trips also have substantial contribution to daily MVPA of young children, which highlights the importance of daily active transport in school- and parental routines.

Determinants of macaques' space use: A test for the ecological constraints model using GPS collars.

As a central topic in Behavioral Ecology, animal space use involves dynamic responses to social and ecological factors. We collared 22 rhesus macaques (Macaca mulatta) from six groups on Neilingding Island, China, and collected 80,625 hourly fixes over a year. Using this high-resolution location data set, we quantified the macaques' space use at the individual level and tested the ecological constraints model while considering various environmental and human interfering factors. As predicted by the ecological constraints model, macaques in larger groups had longer daily path lengths (DPLs) and larger home ranges. We found an inverted U-shape relationship between mean daily temperatures and DPLs, indicating that macaques traveled farther on mild temperature days, while they decreased DPLs when temperatures were too high or too low. Anthropogenic food subsidies were positively correlated to DPLs, while the effect of rainfall was negative. Macaques decreased their DPLs and core areas when more flowers and less leaves were available, suggesting that macaques shifted their space use patterns to adapt to the seasonal differences in food resources. By applying GPS collars on a large number of individuals living on a small island, we gained valuable insights into within-group exploitation competition in wild rhesus macaques.

GRACE Follow-On revealed Bangladesh was flooded early in the 2020 monsoon season due to premature soil saturation.

The overall size and timing of monsoon floods in Bangladesh are challenging to measure. The inundated area is extensive in low-lying Bangladesh, and observations of water storage are key to understanding floods. Laser-ranging instruments on Gravity Recovery and Climate Experiment (GRACE) Follow-On spacecraft detected the peak water storage anomaly of 75 gigatons across Bangladesh in late July 2020. This is in addition to, and three times larger than, the maximum storage anomaly in soil layers during the same period. A flood propagation model suggested that the water mass, as shown in satellite observations, is largely influenced by slow floodplain and groundwater flow processes. Independent global positioning system measurements confirmed the timing and total volume of the flood water estimates. According to land surface models, the soils were saturated a month earlier than the timing of the peak floodplain storage observed by GRACE Follow-On. The cyclone Amphan replenished soils with rainfall just before the monsoon rains started, and consequently, excessive runoff was produced and led to the early onset of the 2020 flooding. This study demonstrated how antecedent soil moisture conditions can influence the magnitude and duration of flooding. Continuous monitoring of storage change from GRACE Follow-On gravity measurements provides important information complementary to river gauges and well levels for enhancing hydrologic flood forecasting models and assisting surface water management.

A data-driven method for identifying the locations of hurricane evacuations from mobile phone location data.

How evacuations are managed can substantially impact the risks faced by affected communities. Having a better understanding of the mobility patterns of evacuees can improve the planning and management of these evacuations. Although mobility patterns during evacuations have traditionally been studied through surveys, mobile phone location data can be used to capture these movements for a greater number of evacuees over a larger geographic area. Several approaches have been used to identify hurricane evacuation patterns from location data; however, each approach relies on researcher judgment to first determine the areas from which evacuations occurred and then identify evacuations by determining when an individual spends a specified number of nights away from home. This approach runs the risk of detecting non-evacuation behaviors (e.g., work trips, vacations, etc.) and incorrectly labeling them as evacuations where none occurred. In this article, we developed a data-driven method to determine which areas experienced evacuations. With this approach, we inferred home locations of mobile phone users, calculated their departure times, and determined if an evacuation may have occurred by comparing the number of departures around the time of the hurricane against historical trends. As a case study, we applied this method to location data from Hurricanes Matthew and Irma to identify areas that experienced evacuations and illustrate how this method can be used to detect changes in departure behavior leading up to and following a hurricane. We validated and examined the inferred homes for representativeness and validated observed evacuation trends against past studies.

Empowerment of Italian general practitioners in depression and suicide prevention: the iFightDepression tool, a game changer of the EAAD-Best project.

General Practitioners (GPs) play a key role in the early detection and management of depression and in preventing suicide risk. They are often the first healthcare professionals that people in crisis contact. However, their effectiveness can be limited by several barriers, including the lack of specific training and appropriate tools.The EAAD-Best project aims to fill these gaps through its iFightDepression tool, an online tool designed to support patients, psychologists, psychiatrists, and GPs in managing depression and preventing suicide. This article examines the implementation of the iFightDepression platform in Italy, assessing its impact on the empowerment of GPs in the fight against depression. Through a qualitative and quantitative analysis of the data collected by the project, the 'unmet need' of GPs' in Italy regarding their specific training in mental health is highlighted.The response of 2,068 Italian GPs in just 7 months after the start of the iFD project is an expression of GPs' engagement to work against depression and for suicide risk prevention.

Association between match-related physical activity profiles and playing positions in different tasks: A data driven approach.

Assessing the intensity characteristics of specific soccer drills (matches, small-side game, and match-based exercises) could help practitioners to plan training sessions by providing the optimal stimulus for every player. In this paper, we propose a data analytics framework to assess the neuromuscular or metabolic characteristics of a soccer-specific exercise in relation with the expected match intensity. GPS data describing the physical tasks' external intensity during an entire season of twenty-eight semi-professional soccer players competing at the fourth Italian division were used in this study. A supervised machine-learning approach was tested in order to detect difference in playing positions in different sport-specific drills. Moreover, a non-supervised machine-learning model was used to profile the match neuromuscular and metabolic characteristics. Players' playing positions during matches and match-based exercises are characterised by specific metabolic and neuromuscular characteristics related to tactical demands, while in the small-side game these differences are not detected. Additionally, our framework permits to evaluate if the match performance request is mirrored during training drills. Practitioners could evaluate the type of stimulus performed by a player in a specific training drill in order to assess if they reflect the matches characteristics of their specific playing position.

Effects of Contextual Variables on Match Load in a Professional Soccer Team Attending to the Different Season Periods.

This study aimed to analyze the effects of contextual variables (i.e., match location and match outcome) and season periods on match load (i.e., internal and external load) in professional Brazilian soccer players. Thirty-six professional players from the same soccer team participated in this study. The season was split into four phases: matches 1-16 (i.e., Phase 1 = P1); matches 17-32 (i.e., Phase 2 = P2); matches 33-48, (i.e., Phase 3 = P3); matches 49-65 (i.e., Phase 4 = P4). Considering match outcome, when the team wins, Cognitive load, Emotional load, and Affective load were significantly higher in away vs. home matches (p < 0.05). Considering season phases, in P3, Mental Fatigue was significantly higher in drawing than in losing matches (p < 0.05). Additionally, considering the match outcome, when the team lost, Total Distance (TD)/min and TD > 19 km·h-1/min were significantly lower in P1 than P2 (p < 0.001), P3 (p < 0.001), and P4 (p < 0.001). These results suggest to strength and conditioning coaches the need to consider the outcome and location of the previous game when planning the week, as well as the phase of the season they are in to reduce fatigue and injury risk.