Escherichia coli CadB is capable of promiscuously transporting muropeptides and contributing to peptidoglycan recycling.

The bacterial peptidoglycan (PG) cell wall is remodeled during growth and division, releasing fragments called muropeptides. Muropeptides can be internalized and reused in a process called PG recycling. Escherichia coli is highly devoted to recycling muropeptides and is known to have at least two transporters, AmpG and OppBCDF, that import them into the cytoplasm. While studying mutants lacking AmpG, we unintentionally isolated mutations that led to the altered expression of a third transporter, CadB. CadB is normally upregulated under acidic pH conditions and is an antiporter for lysine and cadaverine. Here, we explored if CadB was altering PG recycling to assist in the absence of AmpG. Surprisingly, CadB overexpression was able to restore PG recycling when both AmpG and OppBCDF were absent. CadB was found to import freed PG peptides, a subpopulation of muropeptides, through a promiscuous activity. Altogether, our data support that CadB is a third transporter capable of contributing to PG recycling. IMPORTANCE Bacteria produce a rigid mesh cell wall. During growth, the cell wall is remodeled, which releases cell wall fragments. If released into the extracellular environment, cell wall fragments can trigger inflammation by the immune system of a host. Gastrointestinal bacteria, like Escherichia coli, have dedicated pathways to recycle almost all cell wall fragments they produce. E. coli contains two known recycling transporters, AmpG and Opp, that we previously showed are optimized for growth in different environments. Here, we identify that a third transporter, CadB, can also contribute to cell wall recycling. This work expands our understanding of cell wall recycling and highlights the dedication of organisms like E. coli to ensure high recycling in multiple growth environments.

Reputations for treatment of outgroup members can prevent the emergence of political segregation in cooperative networks.

Reputation systems promote cooperation and tie formation in social networks. But how reputations affect cooperation and the evolution of networks is less clear when societies are characterized by fundamental, identity-based, social divisions like those centered on politics in the contemporary U.S. Using a large web-based experiment with participants (N = 1073) embedded in networks where each tie represents the opportunity to play a dyadic iterated prisoners' dilemma, we investigate how cooperation and network segregation varies with whether and how reputation systems track behavior toward members of the opposing political party (outgroup members). As predicted, when participants know others' political affiliation, early cooperation patterns show ingroup favoritism. As a result, networks become segregated based on politics. However, such ingroup favoritism and network-level political segregation is reduced in conditions in which participants know how others behave towards participants from both their own party and participants from the other party. These findings have implications for our understanding of reputation systems in polarized contexts.

Escherichia coli utilizes multiple peptidoglycan recycling permeases with distinct strategies of recycling.

Bacteria produce a structural layer of peptidoglycan (PG) that enforces cell shape, resists turgor pressure, and protects the cell. As bacteria grow and divide, the existing layer of PG is remodeled and PG fragments are released. Enterics such as Escherichia coli go to great lengths to internalize and reutilize PG fragments. E. coli is estimated to break down one-third of its cell wall, yet only loses ~0 to 5% of meso-diaminopimelic acid, a PG-specific amino acid, per generation. Two transporters were identified early on to possibly be the primary permease that facilitates PG fragment recycling, i) AmpG and ii) the Opp ATP binding cassette transporter in conjunction with a PG-specific periplasmic binding protein, MppA. The contribution of each transporter to PG recycling has been debated. Here, we have found that AmpG and MppA/Opp are differentially regulated by carbon source and growth phase. In addition, MppA/Opp is uniquely capable of high-affinity scavenging of muropeptides from growth media, demonstrating that AmpG and MppA/Opp allow for different strategies of recycling PG fragments. Altogether, this work clarifies environmental contexts under which E. coli utilizes distinct permeases for PG recycling and explores how scavenging by MppA/Opp could be beneficial in mixed communities.

Discrimination reduces work effort of those who are disadvantaged and those who are advantaged by it.

Research shows that discrimination is widespread in work organizations, yet we know little about the causal effects of discrimination on employees' work effort. Here we argue that, by decoupling effort from rewards, discrimination reduces the work effort of those who are disadvantaged by discrimination and those advantaged by it. We test these arguments against the results of five experiments designed to model promotion situations in organizations (total N = 1,184). Together, these studies show that when supervised by a manager with a discriminatory preference, both disadvantaged and advantaged workers reduce their work effort relative to a control condition where the manager is not discriminatory. The negative effect of discrimination is larger for those disadvantaged by it. These effects are mediated by employees' beliefs about how strongly work will impact their chances of reward. We then demonstrate that the relatively greater effort of advantaged-versus disadvantaged-workers in discriminatory organizations leads to a self-fulfilling prophecy: when faced with this effort differential, managers (N = 119) who did not have a priori discriminatory attitudes judged the advantaged category as more competent and deserving of workplace advancement than the disadvantaged category. Our results show that even though discrimination reduces all workers' effort, it can ultimately produce outcomes that reify and entrench discriminatory beliefs.

Impact of the cAMP-cAMP Receptor Protein Regulatory Complex on Lipopolysaccharide Modifications and Polymyxin B Resistance in Escherichia coli.

Gram-negative bacteria have a unique cell surface that can be modified to maintain bacterial fitness in diverse environments. A well-defined example is the modification of the lipid A component of lipopolysaccharide (LPS), which promotes resistance to polymyxin antibiotics and antimicrobial peptides. In many organisms, such modifications include the addition of the amine-containing constituents 4-amino-4-deoxy-l-arabinose (l-Ara4N) and phosphoethanolamine (pEtN). Addition of pEtN is catalyzed by EptA, which uses phosphatidylethanolamine (PE) as its substrate donor, resulting in production of diacylglycerol (DAG). DAG is then quickly recycled into glycerophospholipid (GPL) synthesis by the DAG kinase A (DgkA) to produce phosphatidic acid, the major GPL precursor. Previously, we hypothesized that loss of DgkA recycling would be detrimental to the cell when LPS is heavily modified. Instead, we found that DAG accumulation inhibits EptA activity, preventing further degradation of PE, the predominant GPL of the cell. However, DAG inhibition of pEtN addition results in complete loss of polymyxin resistance. Here, we selected for suppressors to find a mechanism of resistance independent of DAG recycling or pEtN modification. Disrupting the gene encoding the adenylate cyclase, cyaA, fully restored antibiotic resistance without restoring DAG recycling or pEtN modification. Supporting this, disruptions of genes that reduce CyaA-derived cAMP formation (e.g., ptsI) or disruption of the cAMP receptor protein, Crp, also restored resistance. We found that loss of the cAMP-CRP regulatory complex was necessary for suppression and that resistance arises from a substantial increase in l-Ara4N-modified LPS, bypassing the need for pEtN modification. IMPORTANCE Gram-negative bacteria can alter the structure of their LPS to promote resistance to cationic antimicrobial peptides, including polymyxin antibiotics. Polymyxins are considered last-resort antibiotics for treatment against multidrug-resistant Gram-negative organisms. Here, we explore how changes in general metabolism and carbon catabolite repression pathways can alter LPS structure and influence polymyxin resistance.

Inequality and cooperation in social networks.

Social networks are fundamental to the broad scale cooperation observed in human populations. But by structuring the flow of benefits from cooperation, networks also create and sustain macro-level inequalities. Here we ask how two aspects of inequality shape the evolution of cooperation in dynamic social networks. Results from a crowdsourced experiment (N = 1080) show that inequality alters the distribution of cooperation within networks such that participants engage in more costly cooperation with their wealthier partners in order to maintain more valuable connections to them. Inequality also influences network dynamics, increasing the tendency for participants to seek wealthier partners, resulting in structural network change. These processes aggregate to alter network structures and produce greater system-level inequality. The findings thus shed critical light on how networks serve as both boon and barrier to macro-level human flourishing.

Ideology shapes how workers perceive and react to workplace discrimination: An experimental study on parenthood discrimination.

Employers use ideologically-tinged rhetoric to justify workplace discrimination. We argue that workers will be less likely to label biased treatment against them as discriminatory when they subscribe to those ideologies as well. We tested this prediction and the consequences of labeling for work attitudes and performance using an experiment that assigned parents to a low-status position in a work group, varying whether the decision invoked biased, ideological assumptions about parenthood. As expected, ideology drove mothers' (but not fathers') labeling. Mothers were less likely to label biased treatment against them as discriminatory when they were conservative and when they subscribed to separate spheres and ideal worker ideologies. Mothers who labeled their treatment as discriminatory had more negative work attitudes than those who did not, but also tended to appeal the decision. Ideology thus shapes whether people label discrimination when it occurs as well as their subsequent work attitudes and justice-seeking behaviors.

Radical flanks of social movements can increase support for moderate factions.

Social movements are critical agents of social change, but are rarely monolithic. Instead, movements are often made up of distinct factions with unique agendas and tactics, and there is little scientific consensus on when these factions may complement-or impede-one another's influence. One central debate concerns whether radical flanks within a movement increase support for more moderate factions within the same movement by making the moderate faction seem more reasonable-or reduce support for moderate factions by making the entire movement seem unreasonable. Results of two online experiments conducted with diverse samples (N = 2,772), including a study of the animal rights movement and a preregistered study of the climate movement, show that the presence of a radical flank increases support for a moderate faction within the same movement. Further, it is the use of radical tactics, such as property destruction or violence, rather than a radical agenda, that drives this effect. Results indicate the effect owes to a contrast effect: Use of radical tactics by one flank led the more moderate faction to appear less radical, even though all characteristics of the moderate faction were held constant. This perception led participants to identify more with and, in turn, express greater support for the more moderate faction. These results suggest that activist groups that employ unpopular tactics can increase support for other groups within the same movement, pointing to a hidden way in which movement factions are complementary, despite pursuing divergent approaches to social change.

Acinetobacter baumannii Can Survive with an Outer Membrane Lacking Lipooligosaccharide Due to Structural Support from Elongasome Peptidoglycan Synthesis.

Gram-negative bacteria resist external stresses due to cell envelope rigidity, which is provided by two membranes and a peptidoglycan layer. The outer membrane (OM) surface contains lipopolysaccharide (LPS; contains O-antigen) or lipooligosaccharide (LOS). LPS/LOS are essential in most Gram-negative bacteria and may contribute to cellular rigidity. Acinetobacter baumannii is a useful tool for testing these hypotheses as it can survive without LOS. Previously, our group found that strains with naturally high levels of penicillin binding protein 1A (PBP1A) could not become LOS deficient unless the gene encoding it was deleted, highlighting the relevance of peptidoglycan biosynthesis and suggesting that high PBP1A levels were toxic during LOS deficiency. Transposon sequencing and follow-up analysis found that axial peptidoglycan synthesis by the elongasome and a peptidoglycan recycling enzyme, ElsL, were vital in LOS-deficient cells. The toxicity of high PBP1A levels during LOS deficiency was clarified to be due to a negative impact on elongasome function. Our data suggest that during LOS deficiency, the strength of the peptidoglycan specifically imparted by elongasome synthesis becomes essential, supporting that the OM and peptidoglycan contribute to cell rigidity. IMPORTANCE Gram-negative bacteria have a multilayered cell envelope with a layer of cross-linked polymers (peptidoglycan) sandwiched between two membranes. Peptidoglycan was long thought to exclusively provide rigidity to the cell providing mechanical strength. Recently, the most outer membrane of the cell was also proposed to contribute to rigidity due to properties of a unique molecule called lipopolysaccharide (LPS). LPS is located on the cell surface in the outer membrane and is typically required for growth. By using Acinetobacter baumannii, a Gram-negative bacterium that can grow without LPS, we found that key features of the peptidoglycan structure also become essential. This finding supports that both the outer membrane and peptidoglycan contribute to cell rigidity.

The Power of Tolerance vs. Unselfishness as a Cultural Determinant of Cooperation.

Cooperation in collective action problems and resource dilemmas is often assumed to depend on the values of the individuals involved, such as their degree of unselfishness and tolerance. Societal differences in cooperation and cooperative norms may therefore result from cultural variation in emphasis on these personal values. Here we draw on several cross-national datasets to examine whether society-level emphasis on unselfishness and tolerance and respect for other people predict how societies vary in cooperation [in a continuous prisoner's dilemma (PD)] and in norms governing cooperation [in a common pool resource dilemma (CPR)]. The results suggest that high levels of cooperation and cooperative norms are promoted specifically by a cultural emphasis on tolerance.

Lipopolysaccharide transport involves long-range coupling between cytoplasmic and periplasmic domains of the LptB2FGC extractor.

The cell surface of the Gram-negative cell envelope contains lipopolysaccharide (LPS) molecules, which form a permeability barrier against hydrophobic antibiotics. The LPS transport (Lpt) machine composed of LptB2FGCADE forms a proteinaceous trans-envelope bridge that allows for the rapid and specific transport of newly synthesized LPS from the inner membrane (IM) to the outer membrane (OM). This transport is powered from the IM by the ATP-binding cassette transporter LptB2FGC. The ATP-driven cycling between closed- and open-dimer states of the ATPase LptB2 is coupled to the extraction of LPS by the transmembrane domains LptFG. However, the mechanism by which LPS moves from a substrate-binding cavity formed by LptFG at the IM to the first component of the periplasmic bridge, the periplasmic ß-jellyroll domain of LptF, is poorly understood. To better understand how LptB2FGC functions in Escherichia coli, we searched for suppressors of a defective LptB variant. We found that defects in LptB2 can be suppressed by both structural modifications to the core oligosaccharide of LPS and changes in various regions of LptFG, including a periplasmic loop in LptF that connects the substrate-binding cavity in LptFG to the periplasmic ß-jellyroll domain of LptF. These novel suppressors suggest that interactions between the core oligosaccharide of LPS and periplasmic regions in the transporter influence the rate of LPS extraction by LptB2FGC. Together, our genetic data reveal a path for the bi-directional coupling between LptB2 and LptFG that extends from the cytoplasm to the entrance to the periplasmic bridge of the transporter.IMPORTANCEGram-negative bacteria are intrinsically resistant to many antibiotics due to the presence of lipopolysaccharide (LPS) at their cell surface. LPS is transported from its site of synthesis at the inner membrane to the outer membrane by the Lpt machine. Lpt proteins form a transporter that spans the entire envelope and is thought to function similarly to a PEZ candy dispenser. This trans-envelope machine is powered by the cytoplasmic LptB ATPase through a poorly understood mechanism. Using genetic analyses in Escherichia coli, we found that LPS transport involves long-ranging bi-directional coupling across cellular compartments between cytoplasmic LptB and periplasmic regions of the Lpt transporter. This knowledge could be exploited in developing antimicrobials that overcome the permeability barrier imposed by LPS.

Restoring Balance to the Outer Membrane: YejM's Role in LPS Regulation.

Gram-negative bacteria produce an asymmetric outer membrane (OM) that is particularly impermeant to many antibiotics and characterized by lipopolysaccharide (LPS) exclusively at the cell surface. LPS biogenesis remains an ideal target for therapeutic intervention, as disruption could kill bacteria or increase sensitivity to existing antibiotics. While it has been known that LPS synthesis is regulated by proteolytic control of LpxC, the enzyme that catalyzes the first committed step of LPS synthesis, it remains unknown which signals direct this regulation. New details have been revealed during study of a cryptic essential inner membrane protein, YejM. Multiple functions have been proposed over the years for YejM, including a controversial hypothesis that it transports cardiolipin from the inner membrane to the OM. Strong evidence now indicates that YejM senses LPS in the periplasm and directs proteolytic regulation. Here, we discuss the standing literature of YejM and highlight exciting new insights into cell envelope maintenance.

The Mla pathway in Acinetobacter baumannii has no demonstrable role in anterograde lipid transport.

The asymmetric outer membrane (OM) of Gram-negative bacteria functions as a selective permeability barrier to the environment. Perturbations to OM lipid asymmetry sensitize the cell to antibiotics. As such, mechanisms involved in lipid asymmetry are fundamental to our understanding of OM lipid homeostasis. One such mechanism, the Maintenance of lipid asymmetry (Mla) pathway has been proposed to extract mislocalized glycerophospholipids from the outer leaflet of the OM and return them to the inner membrane (IM). Work on this pathway in Acinetobacter baumannii support conflicting models for the directionality of the Mla system being retrograde (OM to IM) or anterograde (IM to OM). Here, we show conclusively that A. baumannii mla mutants exhibit no defects in anterograde transport. Furthermore, we identify an allele of the GTPase obgE that is synthetically sick in the absence of Mla; providing another link between cell envelope homeostasis and stringent response.

The robustness of reciprocity: Experimental evidence that each form of reciprocity is robust to the presence of other forms of reciprocity.

Prosocial behavior is paradoxical because it often entails a cost to one's own welfare to benefit others. Theoretical models suggest that prosociality is driven by several forms of reciprocity. Although we know a great deal about how each of these forms operates in isolation, they are rarely isolated in the real world. Rather, the topological features of human social networks are such that people are often confronted with multiple types of reciprocity simultaneously. Does our current understanding of human prosociality break down if we account for the fact that the various forms of reciprocity tend to co-occur in nature? Results of a large experiment show that each basis of human reciprocity is remarkably robust to the presence of other bases. This lends strong support to existing models of prosociality and puts theory and research on firmer ground in explaining the high levels of prosociality observed in human social networks.

LptB-LptF coupling mediates the closure of the substrate-binding cavity in the LptB2 FGC transporter through a rigid-body mechanism to extract LPS.

Lipopolysaccharides (LPS) are essential envelope components in many Gram-negative bacteria and provide intrinsic resistance to antibiotics. LPS molecules are synthesized in the inner membrane and then transported to the cell surface by the LPS transport (Lpt) machinery. In this system, the ATP-binding cassette (ABC) transporter LptB2 FGC extracts LPS from the inner membrane and places it onto a periplasmic protein bridge through a poorly understood mechanism. Here, we show that residue E86 of LptB is essential for coupling the function of this ATPase to that of its partners LptFG, specifically at the step where ATP binding drives the closure of the LptB dimer and the collapse of the LPS-binding cavity in LptFG that moves LPS to the Lpt periplasmic bridge. We also show that defects caused by changing residue E86 are suppressed by mutations altering either LPS structure or transmembrane helices in LptG. Furthermore, these suppressors also fix defects in the coupling helix of LptF, but not of LptG. Together, these results support a transport mechanism in which the ATP-driven movements of LptB and those of the substrate-binding cavity in LptFG are bi-directionally coordinated through the rigid-body coupling, with LptF's coupling helix being important in coordinating cavity collapse with LptB dimerization.

Generosity pays: Selfish people have fewer children and earn less money.

Does selfishness pay in the long term? Previous research has indicated that being prosocial (or otherish) rather than selfish has positive consequences for psychological well-being, physical health, and relationships. Here we instead examine the consequences for individuals' incomes and number of children, as these are the currencies that matter most in theories that emphasize the power of self-interest, namely economics and evolutionary thinking. Drawing on both cross-sectional (Studies 1 and 2) and panel data (Studies 3 and 4), we find that prosocial individuals tend to have more children and higher income than selfish individuals. An additional survey (Study 5) of lay beliefs about how self-interest impacts income and fertility suggests one reason selfish people may persist in their behavior even though it leads to poorer outcomes: people generally expect selfish individuals to have higher incomes. Our findings have implications for lay decisions about the allocation of scarce resources, as well as for economic and evolutionary theories of human behavior. (PsycINFO Database Record (c) 2020 APA, all rights reserved).

Combining Mutations That Inhibit Two Distinct Steps of the ATP Hydrolysis Cycle Restores Wild-Type Function in the Lipopolysaccharide Transporter and Shows that ATP Binding Triggers Transport.

ATP-binding cassette (ABC) transporters constitute a large family of proteins present in all domains of life. They are powered by dynamic ATPases that harness energy from binding and hydrolyzing ATP through a cycle that involves the closing and reopening of their two ATP-binding domains. The LptB2FGC exporter is an essential ABC transporter that assembles lipopolysaccharides (LPS) on the surface of Gram-negative bacteria to form a permeability barrier against many antibiotics. LptB2FGC extracts newly synthesized LPS molecules from the inner membrane and powers their transport across the periplasm and through the outer membrane. How LptB2FGC functions remains poorly understood. Here, we show that the C-terminal domain of the dimeric LptB ATPase is essential for LPS transport in Escherichia coli Specific changes in the C-terminal domain of LptB cause LPS transport defects that can be repaired by intragenic suppressors altering the ATP-binding domains. Surprisingly, we found that each of two lethal changes in the ATP-binding and C-terminal domains of LptB, when present in combined form, suppressed the defects associated with the other to restore LPS transport to wild-type levels both in vivo and in vitro We present biochemical evidence explaining the effect that each of these mutations has on LptB function and how the observed cosuppression results from the opposing lethal effects these changes have on the dimerization state of the LptB ATPase. We therefore propose that these sites modulate the closing and reopening of the LptB dimer, providing insight into how the LptB2FGC transporter cycles to export LPS to the cell surface and how to inhibit this essential envelope biogenesis process.IMPORTANCE Gram-negative bacteria are naturally resistant to many antibiotics because their surface is covered by the glycolipid LPS. Newly synthesized LPS is transported across the cell envelope by the multiprotein Lpt machinery, which includes LptB2FGC, an unusual ABC transporter that extracts LPS from the inner membrane. Like in other ABC transporters, the LptB2FGC transport cycle is driven by the cyclical conformational changes that a cytoplasmic, dimeric ATPase, LptB, undergoes when binding and hydrolyzing ATP. How these conformational changes are controlled in ABC transporters is poorly understood. Here, we identified two lethal changes in LptB that, when combined, remarkably restore wild-type transport function. Biochemical studies revealed that the two changes affect different steps in the transport cycle, having opposing, lethal effects on LptB's dimerization cycle. Our work provides mechanistic details about the LptB2FGC extractor that could be used to develop Lpt inhibitors that would overcome the innate antibiotic resistance of Gram-negative bacteria.

Emerging Roles for NlpE as a Sensor for Lipoprotein Maturation and Transport to the Outer Membrane in Escherichia coli.

Outer membrane biogenesis is a complex process for Gram-negative bacteria as the components are synthesized in the cytoplasm or at the inner membrane and then transported to the outer membrane. Stress pathways monitor and respond to problems encountered in assembling the outer membrane. The two-component system CpxAR was recently reported to be a stress pathway for transport of lipoproteins to the outer membrane, but it was unclear how this stress is sensed. May et al. [K. L. May, K. M. Lehman, A. M. Mitchell, and M. Grabowicz, mBio 10(3):e00618-19, 2019, https://doi.org/10.1128/mBio.00618-19] determined that an outer membrane lipoprotein, NlpE, is the sensor for lipoprotein biogenesis stress. The group demonstrated that CpxAR is activated by the N-terminal domain of NlpE when the lipoprotein accumulates at the inner membrane. Further, this work resolved a previously debated role for NlpE in sensing copper stress; copper was shown to inhibit acylation of lipoproteins, preventing them from being transported to the outer membrane.

Pushing the envelope: LPS modifications and their consequences.

The defining feature of the Gram-negative cell envelope is the presence of two cellular membranes, with the specialized glycolipid lipopolysaccharide (LPS) exclusively found on the surface of the outer membrane. The surface layer of LPS contributes to the stringent permeability properties of the outer membrane, which is particularly resistant to permeation of many toxic compounds, including antibiotics. As a common surface antigen, LPS is recognized by host immune cells, which mount defences to clear pathogenic bacteria. To alter properties of the outer membrane or evade the host immune response, Gram-negative bacteria chemically modify LPS in a wide variety of ways. Here, we review key features and physiological consequences of LPS biogenesis and modifications.

The strength of dynamic ties: The ability to alter some ties promotes cooperation in those that cannot be altered.

Dynamic networks, where ties can be shed and new ties can be formed, promote the evolution of cooperation. Yet, past research has only compared networks where all ties can be severed to those where none can, confounding the benefits of fully dynamic networks with the presence of some dynamic ties within the network. Further, humans do not live in fully dynamic networks. Instead, in real-world networks, some ties are subject to change, while others are difficult to sever. Here, we consider whether and how cooperation evolves in networks containing both static and dynamic ties. We argue and find that the presence of dynamic ties in networks promotes cooperation even in static ties. Consistent with previous work demonstrating that cooperation cascades in networks, our results show that cooperation is enhanced in networks with both tie types because the higher rate of cooperation that occurs following the dynamics process "spills over" to those relations that are more difficult to alter. Thus, our findings demonstrate the critical role that dynamic ties play in promoting cooperation by altering behavioral outcomes even in non-dynamic relations.