Sequence variation among homologous proteins can shed light on their function and ancestry. In this study, we analyze variation at catalytic residues among MCR (mobile colistin resistance) proteins, which confer resistance to the last resort antibiotic, colistin, in gram-negative bacteria. We show that not all naturally occurring variants at a lipid A-binding residue, Ser284, are tolerated in MCR-1. In particular, the substitution of Ser284 with Asp, found naturally in MCR-5, resulted in diminished colistin resistance. Using phylogenetic analyses and structure predictions we trace back variation at this site among MCRs to their ancestors, i.e. EptA phosphoethanolamine transferases that are encoded by diverse bacterial genomes. Mutational studies and AlphaFold-based structural modeling revealed that the functional importance of position 284 varies between phylogenetically distant MCRs, i.e. MCR-1 and MCR-5. Despite a high degree of similarity among their catalytic domains, inter-domain interactions were not conserved between MCR-1 and MCR-5 due to their different ancestries, providing a mechanistic basis behind the different phenotypes of similar mutations at position 284. Our study thus uncovers subtle differences in the organization of domains among MCR proteins that can lead to substantial differences in their catalytic properties and mutational tolerances.

Jonas Kellermeyer responds in Round 2 to the various critiques of the Artistic Research working group, see Round 1. Subsequently, Till Bödeker/Peter Tepe reply to Kellermeyer’s response.

Cell motility is crucial for many biological processes including morphogenesis, wound healing, and cancer invasion. The WAVE regulatory complex (WRC) is a central Arp2/3 regulator driving cell motility downstream of activation by Rac GTPase. CYFIP-related Rac1 interactor (CYRI) proteins are thought to compete with WRC for interaction with Rac1 in a feedback loop regulating lamellipodia dynamics. However, the physiological role of CYRI proteins in vivo in healthy tissues is unclear. Here, we used Drosophila as a model system to study CYRI function at the cellular and organismal levels. We found that CYRI is not only a potent WRC regulator in single macrophages that controls lamellipodial spreading but also identified CYRI as a molecular brake on the Rac-WRC-Arp2/3 pathway to slow down epidermal wound healing. In addition, we found that CYRI limits invasive border cell migration by controlling cluster cohesion and migration. Thus, our data highlight CYRI as an important regulator of cellular and epithelial tissue dynamics conserved across species.

Cells maintain homeostasis via dynamic regulation of stress response pathways. Stress pathways transiently induce response regulons via negative feedback loops, but the extent to which individual genes provide feedback has not been comprehensively measured for any pathway. Here, we disrupted the induction of each gene in the Saccharomyces cerevisiae heat shock response (HSR) and quantified cell growth and HSR dynamics following heat shock. The screen revealed a core feedback loop governing the expression of the chaperone Hsp70 reinforced by an auxiliary feedback loop controlling Hsp70 subcellular localization. Mathematical modeling and live imaging demonstrated that multiple HSR targets converge to promote Hsp70 nuclear localization via its release from cytosolic condensates. Following ethanol stress, a distinct set of factors similarly converged on Hsp70, suggesting that nonredundant subsets of the HSR regulon confer feedback under different conditions. Flexible spatiotemporal feedback loops may broadly organize stress response regulons and expand their adaptive capacity.

Ecovillages are settlements that aim to combine human well-being and environmental sustainability. They serve as sites of social learning for sustainable consumption practices and may inspire lifestyle changes in mainstream society. However, most ecovillages fail in the first few years of establishment. Through an in-depth study of a Norwegian ecovillage’s efforts at expansion, this article explores the factors that adversely affected the well-being of ecovillage residents and threatened the community’s survival. The study applies mixed methods combining needs-based workshops, a well-being survey, and in-depth interviews. The transformation of the ecovillage into a larger settlement with modern houses was driven by a developer and with minimal involvement of residents. The resulting divergence in visions and values, coupled with structural problems of the homes, led to protracted conflicts, and resulted in the developer’s bankruptcy. The study identifies strong social institutions and affordable infrastructures as crucial elements for both well-being and the success of ecovillages.

Penelitian ini bertujuan untuk mengetahui dan menganalisis seberapa besar pengaruh terpaan iklan melalui daya tarik iklan terhadap sikap audiens pada iklan (studi kasus pada audiens iklan Pantene Miracles di Kota Depok). Tujuan penelitian ini adalah untuk menganalisis seberapa besar pengaruh terpaan iklan terhadap sikap audiens pada iklan melalui daya tarik iklan Pantene Miracles.Metode penelitian ini adalah kuantitatif dengan pendekatan penelitian eksplanasi. Metode pengambilan sampel dalam penelitian ini menggunakan metode nonprobability sampling dan teknik purposive sampling dengan responden sebanyak 100 responden. Kemudian diuji dengan menggunakan PLS (Partial Least Square). Pengujian instrumen menggunakan uji validitas dan uji reliabilitas. Sedangkan untuk metode analisis data menggunakan uji validitas diskriminan, uji validitas konvergen, uji reliabilitas, dan uji hipotesis. Hasil penelitian ini menunjukkan bahwa: (a) Terpaan iklan (X) berpengaruh positif sebesar 45,8% terhadap sikap audiens pada iklan (Y); (b) Terpaan iklan (X) berpengaruh positif sebesar 64,1% terhadap daya tarik iklan (Z); (c) Daya tarik iklan (Z) berpengaruh positif sebesar 46,2% terhadap sikap audiens pada iklan (Y); (d) Terpaan iklan (X) berpengaruh positif sebesar 29,6% terhadap sikap audiens pada iklan (Y) melalui daya tarik iklan (Z).Hasil uji hipotesis pada penelitian ini menunjukkan bahwa terdapat pengaruh positif dan signifikan antara variabel terpaan iklan, daya tarik iklan, dan sikap audiens pada iklan sehingga semua Ha diterima dan H0 ditolak.

Second-generation practice theoretical research on sustainable consumption has largely overlooked the role of environmental ethics. This is likely due to associations of “ethics” with culture and symbolic understandings of consumption. This article investigates how some environmentally aware and engaged consumers actively try to cultivate ethical and sustainable practices in order to develop ways of living more sustainably. By combining a practice theoretical framework with Michel Foucault’s work on ethical self-cultivation, I combine the ethical – understood as emerging in specific places and time as part of cultural formations – with processes of embodiment and ethical reflexivity to understand how environmental ethical reflexivity can play a role in the sustainable change of practices. The article argues that changing consumption practices based on changed environmental ethics requires an ongoing reflexive and bodily process of training the bodymind to embody skills and mold sensory, affective, and perceptual dispositions. This is a processual change requiring time and effort, in which research participants nevertheless experience meaning and hope as they are able to create sustainable change in their own lives and, over time and to some degree, live up to their ethical ideals. The analysis is based on ethnographic fieldwork conducted in two locations using audio-visual methods in participant observation and interviews with research participants who are actively engaged in changing their everyday practices.

Mutations in VPS13B, a member of a protein family implicated in bulk lipid transport between adjacent membranes, cause Cohen syndrome. VPS13B is known to be concentrated in the Golgi complex, but its precise location within this organelle and thus the site(s) where it achieves lipid transport remains unclear. Here, we show that VPS13B is localized at the interface between proximal and distal Golgi subcompartments and that Golgi complex reformation after Brefeldin A (BFA)-induced disruption is delayed in VPS13B KO cells. This delay is phenocopied by the loss of FAM177A1, a Golgi complex protein of unknown function reported to be a VPS13B interactor and whose mutations also result in a developmental disorder. In zebrafish, the vps13b ortholog, not previously annotated in this organism, genetically interacts with fam177a1. Collectively, these findings raise the possibility that bulk lipid transport by VPS13B may play a role in the dynamics of Golgi membranes and that VPS13B may be assisted in this function by FAM177A1.