Clinical Education Data Management: Current Landscape in Entry-Level Physical Therapist Education.

BACKGROUND: Data management (DM) systems represent an opportunity for innovation in education and data-driven decision-making (DDDM) in allied health education. Understanding clinical education (CE) DM systems in entry-level physical therapy (PT) education programs could provide valuable insight into structure and operation and may represent opportunities to address CE challenges. The purpose of this study is to describe how PT programs are using CE DM systems to inform recommendations for CE DM and support knowledge sharing and DDDM. SUBJECTS: CE faculty and administrators were recruited from entry-level PT education programs to participate in a cross-sectional survey. METHODS: The authors designed a novel survey which included demographics and use of CE DM systems. Descriptive statistics and content analysis of narrative data were used to examine responses. RESULTS: The survey was distributed to 220 academic PT programs in June 2021 with 111 respondents (50% response rate). Respondents use multiple systems to complete CE tasks (e.g., placement process, on-boarding, agreement tracking, as a CE site database). Forty-three percent (n=47) use one system, 76% (n=35) of those use the same Software as a Service vendor. Eighty-six percent (n=96) are satisfied with their current CE DM system. Respondents enter data related to CE site information, CE environment, length of the CE experience, and accreditation-required clinical instructor information. Ninety-four percent (n=93) and 70% (n=70) extract data to make decisions about the placement process and curriculum, respectively. CONCLUSION: While variability across CE DM systems presents a challenge, survey respondents indicated common practices related to functionality, data entry, and extraction. Clinical education DM systems house critical data to address challenges in CE. Strategies to improve accessibility and use of this data to support DDDM should be explored.

Defining Biological and Biochemical Functions of Noncanonical SET Domain Proteins.

Within the SET domain superfamily of lysine methyltransferases, there is a well-conserved subfamily, frequently referred to as the Set3 SET domain subfamily, which contain noncanonical SET domains carrying divergent amino acid sequences. These proteins are implicated in diverse biological processes including stress responses, cell differentiation, and development, and their disruption is linked to diseases including cancer and neurodevelopmental disorders. Interestingly, biochemical and structural analysis indicates that they do not possess catalytic methyltransferase activity. At the molecular level, Set3 SET domain proteins appear to play critical roles in the regulation of gene expression, particularly repression and heterochromatin maintenance, and in some cases, via scaffolding other histone modifying activities at chromatin. Here, we explore the common and unique functions among Set3 SET domain subfamily proteins and analyze what is known about the specific contribution of the conserved SET domain to functional roles of these proteins, as well as propose areas of investigation to improve understanding of this important, noncanonical subfamily of proteins.

The SMYD3-MAP3K2 signaling axis promotes tumor aggressiveness and metastasis in prostate cancer.

Aberrant activation of Ras/Raf/mitogen-activated protein kinase (MAPK) signaling is frequently linked to metastatic prostate cancer (PCa); therefore, the characterization of modulators of this pathway is critical for defining therapeutic vulnerabilities for metastatic PCa. The lysine methyltransferase SET and MYND domain 3 (SMYD3) methylates MAPK kinase kinase 2 (MAP3K2) in some cancers, causing enhanced activation of MAPK signaling. In PCa, SMYD3 is frequently overexpressed and associated with disease severity; however, its molecular function in promoting tumorigenesis has not been defined. We demonstrate that SMYD3 critically regulates tumor-associated phenotypes via its methyltransferase activity in PCa cells and mouse xenograft models. SMYD3-dependent methylation of MAP3K2 promotes epithelial-mesenchymal transition associated behaviors by altering the abundance of the intermediate filament vimentin. Furthermore, activation of the SMYD3-MAP3K2 signaling axis supports a positive feedback loop continually promoting high levels of SMYD3. Our data provide insight into signaling pathways involved in metastatic PCa and enhance understanding of mechanistic functions for SMYD3 to reveal potential therapeutic opportunities for PCa.

Restriction of Arginine Induces Antibiotic Tolerance in Staphylococcus aureus.

Staphylococcus aureus is responsible for a substantial number of invasive infections globally each year. These infections are problematic because they are frequently recalcitrant to antibiotic treatment, particularly when they are caused by Methicillin-Resistant Staphylococcus aureus (MRSA). Antibiotic tolerance, the ability for bacteria to persist despite normally lethal doses of antibiotics, is responsible for most antibiotic treatment failure in MRSA infections. To understand how antibiotic tolerance is induced, S. aureus biofilms exposed to multiple anti-MRSA antibiotics (vancomycin, ceftaroline, delafloxacin, and linezolid) were examined using both quantitative proteomics and transposon sequencing. These screens indicated that arginine metabolism is involved in antibiotic tolerance within a biofilm and led to the hypothesis that depletion of arginine within S. aureus communities can induce antibiotic tolerance. Consistent with this hypothesis, inactivation of argH, the final gene in the arginine synthesis pathway, induces antibiotic tolerance under conditions in which the parental strain is susceptible to antibiotics. Arginine restriction was found to induce antibiotic tolerance via inhibition of protein synthesis. Finally, although S. aureus fitness in a mouse skin infection model is decreased in an argH mutant, its ability to survive in vivo during antibiotic treatment with vancomycin is enhanced, highlighting the relationship between arginine metabolism and antibiotic tolerance during S. aureus infection. Uncovering this link between arginine metabolism and antibiotic tolerance has the potential to open new therapeutic avenues targeting previously recalcitrant S. aureus infections.

Hyperglycemia Increases Severity of Staphylococcus aureus Osteomyelitis and Influences Bacterial Genes Required for Survival in Bone.

Hyperglycemia, or elevated blood glucose, renders individuals more prone to developing severe Staphylococcus aureus infections. S. aureus is the most common etiological agent of musculoskeletal infection, which is a common manifestation of disease in hyperglycemic patients. However, the mechanisms by which S. aureus causes severe musculoskeletal infection during hyperglycemia are incompletely characterized. To examine the influence of hyperglycemia on S. aureus virulence during invasive infection, we used a murine model of osteomyelitis and induced hyperglycemia with streptozotocin. We discovered that hyperglycemic mice exhibited increased bacterial burdens in bone and enhanced dissemination compared to control mice. Furthermore, infected hyperglycemic mice sustained increased bone destruction relative to euglycemic controls, suggesting that hyperglycemia exacerbates infection-associated bone loss. To identify genes contributing to S. aureus pathogenesis during osteomyelitis in hyperglycemic animals relative to euglycemic controls, we used transposon sequencing (TnSeq). We identified 71 genes uniquely essential for S. aureus survival in osteomyelitis in hyperglycemic mice and another 61 mutants with compromised fitness. Among the genes essential for S. aureus survival in hyperglycemic mice was the gene encoding superoxide dismutase A (sodA), one of two S. aureus superoxide dismutases involved in detoxifying reactive oxygen species (ROS). We determined that a sodA mutant exhibits attenuated survival in vitro in high glucose and in vivo during osteomyelitis in hyperglycemic mice. SodA therefore plays an important role during growth in high glucose and promotes S. aureus survival in bone. Collectively, these studies demonstrate that hyperglycemia increases the severity of osteomyelitis and identify genes contributing to S. aureus survival during hyperglycemic infection.

Set1 regulates telomere function via H3K4 methylation-dependent and -independent pathways and calibrates the abundance of telomere maintenance factors.

Set1 is an H3K4 methyltransferase that comprises the catalytic subunit of the COMPASS complex and has been implicated in transcription, DNA repair, cell cycle control, and numerous other genomic functions. Set1 also promotes proper telomere maintenance, as cells lacking Set1 have short telomeres and disrupted subtelomeric gene repression; however, the precise role for Set1 in these processes has not been fully defined. In this study, we have tested mutants of Set1 and the COMPASS complex that differentially alter H3K4 methylation status, and we have attempted to separate catalytic and noncatalytic functions of Set1. Our data reveal that Set1-dependent subtelomeric gene repression relies on its catalytic activity toward H3K4, whereas telomere length is regulated by Set1 catalytic activity but likely independent of the H3K4 substrate. Furthermore, we uncover a role for Set1 in calibrating the abundance of critical telomere maintenance proteins, including components of the telomerase holoenzyme and members of the telomere capping CST (Cdc13-Stn1-Ten1) complex, through both transcriptional and posttranscriptional pathways. Altogether, our data provide new insights into the H3K4 methylation-dependent and -independent roles for Set1 in telomere maintenance in yeast and shed light on possible roles for Set1-related methyltransferases in other systems.

The addition of body weight supported treadmill training to manual therapy and exercise in the management of Hip osteoarthritis: A case series.

BACKGROUND AND PURPOSE: Patients with mild-to-moderate hip OA can present with pain, a decline in function, altered gait mechanics, and pain with ambulation. Body weight supported treadmill training (BWSTT) has been utilized for patients with total hip arthroplasty, hip fracture, and lumbar spinal stenosis. The purpose of this case series was to report the outcomes of patients with hip OA that received guideline adherent physical therapy care with the addition of BWSTT. Our aim was to assess changes in pain, disability, and physical performance. CASE DESCRIPTIONS: Seven patients participated in eight 1-h treatment sessions consisting of: manual therapy, therapeutic exercise, and BWSTT. Pre- and post-treatment outcome measures included: average pain rating via the Numeric Pain Rating Scale (NPRS), Western Ontario and McMasters Universities Osteoarthritis Index (WOMAC), and the Global Rate of Change (GROC). Physical performance measures included: 6-minute walk test (6MWT), stair climbing test, and 5 time sit-stand. OUTCOMES: The mean improvement in NPRS score for all subjects was 2.9 points. Mean improvement on the WOMAC was 18.5 + 24.8 pts, and the mean GROC score was +5.0 indicating a rating of "quite a bit better." The mean increase in 6MWT distance was 60.5 + 80.1 meters (median 39 m, range -3 to 230). CONCLUSIONS: Though patients participating in a multimodal rehabilitation approach including manual therapy, exercise, and BWSTT demonstrated clinically important improvements in pain and function, these changes may not correspond with gait endurance improvements in the short term.

Hospitalization Outcomes Among Patients With COVID-19 Undergoing Remote Monitoring.

Importance: Health care systems have implemented remote patient monitoring (RPM) programs to manage patients with COVID-19 at home, but the associations between participation and outcomes or resource utilization are unclear. Objective: To assess whether an RPM program for COVID-19 is associated with lower or higher likelihood of hospitalization and whether patients who are admitted present earlier or later for hospital care. Design, Setting, and Participants: This retrospective, observational, cohort study of RPM was performed at Froedtert & Medical College of Wisconsin Health Network, an academic health system in southeastern Wisconsin. Participants included patients with internal primary care physicians and a positive SARS-CoV-2 test in the ambulatory setting between March 30, 2020, and December 15, 2020. Data analysis was performed from February 15, 2021, to February 2, 2022. Exposures: Activation of RPM program. Main Outcomes and Measures: Hospitalizations within 2 to 14 days of a positive test. Inverse propensity score weighting was used to account for differences between groups. Sensitivity analyses were performed looking at usage of the RPM among patients who activated the program. Results: A total of 10 660 COVID-19-positive ambulatory patients were eligible, and 9378 (88.0%) had email or mobile numbers on file and were invited into the RPM program; the mean (SD) age was 46.9 (16.3) years and 5448 patients (58.1%) were women. Patients who activated monitoring (5364 patients [57.2%]) had a mean (SD) of 35.3 (33.0) check-ins and a mean (SD) of 1.27 (2.79) (median [IQR], 0 [0-1]) free-text comments. A total of 878 patients (16.4%) experienced at least 1 alert; 128 of 5364 activated patients (2.4%) and 158 of 4014 inactivated patients (3.9%) were hospitalized (χ21 = 18.65; P < .001). In weighted regression analysis, activation of RPM was associated with a lower odds of hospitalization (odds ratio, 0.68; 95% CI, 0.54-0.86; P = .001) adjusted for demographics, comorbidities, and time period. Monitored patients had a longer mean (SD) time between test and hospitalization (6.67 [3.21] days vs 5.24 [3.03] days), a shorter length of stay (4.44 [4.43] days vs 7.14 [8.63] days), and less intensive care use (15 patients [0.3%] vs 44 patients [1.1%]). Conclusions and Relevance: These findings suggest that activation of an RPM program is associated with lower hospitalization, intensive care use, and length of stay among patients with COVID-19.

Bacterial hydrophilins promote pathogen desiccation tolerance.

Acinetobacter baumannii is a leading cause of hospital-acquired infections, where outbreaks are driven by its ability to persist on surfaces in a desiccated state. Here, we show that A. baumannii causes more virulent pneumonia following desiccation and profile the genetic requirements for desiccation. We find that desiccation tolerance is enhanced upon the disruption of Lon protease, which targets unfolded and aggregated proteins for degradation. Notably, two bacterial hydrophilins, DtpA and DtpB, are transcriptionally upregulated in Δlon via the two-component regulator, BfmR. These proteins, both hydrophilic and intrinsically disordered, promote desiccation tolerance in A. baumannii. Additionally, recombinant DtpA protects purified enzymes from inactivation and improves the desiccation tolerance of a probiotic bacterium when heterologously expressed. These results demonstrate a connection between environmental persistence and pathogenicity in A. baumannii, provide insight into the mechanisms of extreme desiccation tolerance, and reveal potential applications for bacterial hydrophilins in the preservation of protein- and live bacteria-based pharmaceuticals.

Immunoaffinity purification of endogenous proteins from S. cerevisiae for post-translational modification and protein interaction analysis.

Protein regulation by post-translational modifications and protein-protein interactions is critical to controlling molecular pathways. Here, we describe an immunoaffinity purification approach in Saccharomyces cerevisiae. The protocol uses an endogenously-expressed epitope-tagged protein and can be applied to the identification of post-translational modifications or protein binding partners. The lysine methyltransferase Set5 is used as an example here to purify phosphorylated Set5 and identify phosphosites; however, this approach can be applied to a diverse set of proteins in yeast. For complete details on the use and execution of this protocol, please refer to Jaiswal et al. (2020).

Set4 regulates stress response genes and coordinates histone deacetylases within yeast subtelomeres.

The yeast chromatin protein Set4 is a member of the Set3-subfamily of SET domain proteins which play critical roles in the regulation of gene expression in diverse developmental and environmental contexts. We previously reported that Set4 promotes survival during oxidative stress and regulates expression of stress response genes via stress-dependent chromatin localization. In this study, global gene expression analysis and investigation of histone modification status identified a role for Set4 in maintaining gene repressive mechanisms within yeast subtelomeres under both normal and stress conditions. We show that Set4 works in a partially overlapping pathway to the SIR complex and the histone deacetylase Rpd3 to maintain proper levels of histone acetylation and expression of stress response genes encoded in subtelomeres. This role for Set4 is particularly critical for cells under hypoxic conditions, where the loss of Set4 decreases cell fitness and cell wall integrity. These findings uncover a new regulator of subtelomeric chromatin that is key to stress defense pathways and demonstrate a function for Set4 in regulating repressive, heterochromatin-like environments.

Migrants and Service Providers' Perspectives of Barriers to Accessing Mental Health Services in South Australia: A Case of African Migrants with a Refugee Background in South Australia.

International mobility has increased steadily in recent times, bringing along a myriad of health, social and health system challenges to migrants themselves and the host nations. Mental health issues have been identified as a significant problem among migrants, with poor accessibility and underutilisation of the available mental health services (MHSs) repeatedly reported, including in Australia. Using a qualitative inquiry and one-on-one in-depth interviews, this study explored perspectives of African migrants and service providers on barriers to accessing MHSs among African migrants in South Australia. The data collection took place during the COVID-19 pandemic with lockdown and other measures to combat the pandemic restricting face to face meetings with potential participants. Online platforms including Zoom and/or WhatsApp video calls were used to interview 20 African migrants and 10 service providers. Participants were recruited from community groups and/or associations, and organisations providing services for migrants and/or refugees in South Australia using the snowball sampling technique. Thematic framework analysis was used to guide the data analysis. Key themes centred on personal factors (health literacy including knowledge and the understanding of the health system, and poor financial condition), structural factors related to difficulties in navigating the complexity of the health system and a lack of culturally aware service provision, sociocultural and religious factors, mental health stigma and discrimination. The findings provide an insight into the experiences of African migrants of service provision to them and offer suggestions on how to improve these migrants' mental health outcomes in Australia. Overcoming barriers to accessing mental health services would need a wide range of strategies including education on mental health, recognising variations in cultures for effective service provision, and addressing mental health stigma and discrimination which strongly deter service access by these migrants. These strategies will facilitate help-seeking behaviours as well as effective provision of culturally safe MHSs and improvement in access to MHSs among African migrants.

Adaptive local false discovery rate procedures for highly spiky data and their application RNA sequencing data of yeast SET4 deletion mutants.

Chromatin dynamics are central to the regulation of gene expression and genome stability. In order to improve understanding of the factors regulating chromatin dynamics, the genes encoding these factors are deleted and the differential gene expression profiles are determined using approaches such as RNA sequencing. Here, we analyzed a gene expression dataset aimed at uncovering the function of the relatively uncharacterized chromatin regulator, Set4, in the model system Saccharomyces cerevisiae (budding yeast). The main theme of this paper focuses on identifying the highly differentially expressed genes in cells deleted for Set4 (referred to as Set4 Δ mutant dataset) compared to the wild-type yeast cells. The Set4 Δ mutant data produce a spiky distribution on the log-fold changes of their expressions, and it is reasonably assumed that genes which are not highly differentially expressed come from a mixture of two normal distributions. We propose an adaptive local false discovery rate (FDR) procedure, which estimates the null distribution of the log-fold changes empirically. We numerically show that, unlike existing approaches, our proposed method controls FDR at the aimed level (0.05) and also has competitive power in finding differentially expressed genes. Finally, we apply our procedure to analyzing the Set4 Δ mutant dataset.

Personal, proxy, and collective food agency among early adolescents.

Early adolescence is a critical time for health behavior development because agency increases during the transition from childhood to adolescence. This qualitative study sought to identify how early adolescent participants described food-related agency. One-on-one interviews were conducted with 30 early adolescents (10-13 years). Data analysis was guided by Bandura's three modes of agency: personal, proxy, and collective. Results suggest participants' food behaviors were informed by a growing knowledge about nutrition, household food rules, and school food environments. Participants described different modes of agency in four areas - grocery shopping, cooking, consumption decisions, and nutrition information seeking - with varying degrees of agency in each area. Understanding how each of the three modes operate and the interplay between them can information future research aimed at improving the nutrition behaviors of early adolescents.

Addressing depression and behavioral health needs through a digital program at scale.

Depression and anxiety disorders are prevalent mental health conditions; yet they are often unrecognized, under-addressed and/or under-treated, and specialty treatment for these conditions is oftentimes difficult to access. By acting either as a bridge to therapy or as a form of therapy, digital tools, such as those that provide internet-based cognitive behavioral therapy (iCBT), may help clinicians support their patients' mental health needs. At one academic health system, a digital mental health program was deployed in primary care and outpatient behavioral health programs to help patients meet needs identified through screening or clinical visits. Over the first two years of operation, 138 clinicians (40% of eligible clinicians) prescribed the program to 2,228 unique patients, from which 1,117 (48.9%) enrolled. Patients who enrolled tended to be younger and healthier than non-enrollees. On average, enrolled patients spent 114.6 minutes within the iCBT program. Clinical improvement was assessed using pre- and post PHQ-9 and GAD-7 scores for depression and anxiety, respectively. Pre/Post scores were compared using Wilcoxon Rank Sum test. Patients with at least moderate depression had an average 23% reduction in PHQ-9 scores (median change -3(interquartile range 7), p<0.001) and those with at least moderate anxiety had a 26% reduction in GAD-7 scores (-4(7), p<0.001). Improvements were clinically and statistically significant. Future steps include performing a cost analysis to understand whether models utilizing iCBT are net cost-saving for health systems.

Early adolescent food routines: A photo-elicitation study.

Early adolescence (ages 10-14) encompasses a critical transition period in which food and nutrition decisions are shifting in important ways. Food routines are food-based activities that repeat across days, weeks, seasons, or lives. Examining routines can provide insight into how individuals are influenced in food choices. The objective of this study was to describe current influences on and experiences with food routines during early adolescence. In-depth interviews, using a photo-elicitation approach, were conducted with 30 participants (16 females; 14 males) in the United States. Participants took photos that were then used during the interview to describe food-related decisions and influences. Interviews were audio recorded and transcribed verbatim. Analysis was guided by a grounded theory approach to identify emergent themes related to routines and resulted in the development of a conceptual model for early adolescent food routines. Participants identified a wide range of routines and three main themes emerged: family, settings, and meals/foods consumed. Some had highly established routines throughout the week, while others described routines only for certain meals or days. Several participants described increased control or the ability to modify routines around some eating episodes such as snacks, lunches, and weekend breakfasts. Findings revealed how participants viewed eating routines and provided information about food-and nutrition-related behaviors that can inform future research and practice. Early adolescents appear to have complex food routines influenced by structures and different amounts of control.

Psychological trauma and access to primary healthcare for people from refugee and asylum-seeker backgrounds: a mixed methods systematic review.

BACKGROUND: Several reviews have found that psychological trauma affects access to health care services, including mental health care, in the general population. People from refugee and asylum seeker backgrounds are more likely to have a mental illness than the general population, and experience a broad range of barriers and facilitators to service access. However, to date there has been no comprehensive consideration of the potential effect of psychological trauma on access to primary health care within this population. METHODS: This paper provides a mixed-methods systematic review of literature which included any consideration of the relationship between psychological trauma and access to primary health care. A systematic search of Medline, PsychInfo, Scopus, Web of Science, Embase, CINAHL and Cochrane Library was conducted. Study eligibility criteria were empirical, peer-reviewed studies that considered the relationship between psychological trauma and access to, or use of, primary healthcare in resettlement countries for refugees (including asylum seekers). Papers were required to be written in English and published between 1998 and August 2019. Quality was assessed using the Multi-Methods Appraisal Tool. The search identified a total of 14 eligible studies (11 quantitative and 3 qualitative) which had explored this relationship in refugee and asylum seeker populations. RESULTS: Overall, synthesis of findings indicated variable results with respect to the impact of psychological trauma on service access. Specifically, the review found that while rates of psychological trauma were high. Key themes were that while general health care access was comparable or greater than the general population, rates of mental healthcare specifically were low. In addition, included papers identified a range of barriers to service access-particularly somatisation, stigma and healthcare provide knowledge about psychological trauma. CONCLUSIONS: While there is a critical need for more research in this area, the study points to several key recommendations including training of general practitioners in relation to psychological trauma, ensuring culturally responsive services, and the use of interpreters. Finally, due to the levels of somatisation found in some studies, ensuring general practitioners understand the somatic element of psychological trauma-particularly within some groups of people from refugee backgrounds-is important.

Using Yeast to Define the Regulatory Role of Protein Lysine Methylation.

The post-translational modifications (PTM) of proteins are crucial for cells to survive under diverse environmental conditions and to respond to stimuli. PTMs are known to govern a broad array of cellular processes including signal transduction and chromatin regulation. The PTM lysine methylation has been extensively studied within the context of chromatin and the epigenetic regulation of the genome. However, it has also emerged as a critical regulator of non-histone proteins important for signal transduction pathways. While the number of known non-histone protein methylation events is increasing, the molecular functions of many of these modifications are not yet known. Proteomic studies of the model system Saccharomyces cerevisiae suggest lysine methylation may regulate a diversity of pathways including transcription, RNA processing, translation, and signal transduction cascades. However, there has still been relatively little investigation of lysine methylation as a broad cellular regulator beyond chromatin and transcription. Here, we outline our current state of understanding of non-histone protein methylation in yeast and propose ways in which the yeast system can be leveraged to develop a much more complete picture of molecular mechanisms through which lysine methylation regulates cellular functions.

The Manganese-Responsive Transcriptional Regulator MumR Protects Acinetobacter baumannii from Oxidative Stress.

Acinetobacter baumannii is an emerging opportunistic pathogen that primarily infects critically ill patients in nosocomial settings. Because of its rapid acquisition of antibiotic resistance, infections caused by A. baumannii have become extremely difficult to treat, underlying the importance of identifying new antimicrobial targets for this pathogen. Manganese (Mn) is an essential nutrient metal required for a number of bacterial processes, including the response to oxidative stress. Here, we show that exogenous Mn can restore A. baumannii viability in the presence of reactive oxygen species (ROS). This restoration is not dependent on the high-affinity Nramp family Mn transporter, MumT, as a ΔmumT mutant is no more sensitive to hydrogen peroxide (H2O2) killing than wild-type A. baumannii However, mumR, which encodes the transcriptional regulator of mumT, is critical for growth and survival in the presence of H2O2, suggesting that MumR regulates additional genes that contribute to H2O2 resistance. RNA sequencing revealed a role for mumR in regulating the activity of a number of metabolic pathways, including two pathways, phenylacetate and gamma-aminobutyric acid catabolism, which were found to be important for resisting killing by H2O2 Finally, ΔmumR exhibited reduced fitness in a murine model of pneumonia, indicating that MumR-regulated gene products are crucial for protection against the host immune response. In summary, these results suggest that MumR facilitates resistance to the host immune response by activating a transcriptional program that is critical for surviving both Mn starvation and oxidative stress.

Function of the MYND Domain and C-Terminal Region in Regulating the Subcellular Localization and Catalytic Activity of the SMYD Family Lysine Methyltransferase Set5.

SMYD lysine methyltransferases target histones and nonhistone proteins for methylation and are critical regulators of muscle development and implicated in neoplastic transformation. They are characterized by a split catalytic SET domain and an intervening MYND zinc finger domain, as well as an extended C-terminal domain. Saccharomyces cerevisiae contains two SMYD proteins, Set5 and Set6, which share structural elements with the mammalian SMYD enzymes. Set5 is a histone H4 lysine 5, 8, and 12 methyltransferase, implicated in the regulation of stress responses and genome stability. While the SMYD proteins have diverse roles in cells, there are many gaps in our understanding of how these enzymes are regulated. Here, we performed mutational analysis of Set5, combined with phosphoproteomics, to identify regulatory mechanisms for its enzymatic activity and subcellular localization. Our results indicate that the MYND domain promotes Set5 chromatin association in cells and is required for its role in repressing subtelomeric genes. Phosphoproteomics revealed extensive phosphorylation of Set5, and phosphomimetic mutations enhance Set5 catalytic activity but diminish its ability to interact with chromatin in cells. These studies uncover multiple regions within Set5 that regulate its localization and activity and highlight potential avenues for understanding mechanisms controlling the diverse roles of SMYD enzymes.