Hi-C Contacts Encode Heterogeneity in Sub-diffusive Motion of E. coli Chromosomal Loci

Underneath its apparently simple architecture, the circular chromosome of E. coli is known for displaying complex dynamics in its cytoplasm. Recent experiments have hinted at an inherently heterogeneous dynamics of chromosomal loci, the origin of which has largely been elusive. In this regard, here we investigate the loci dynamics of E. coli chromosome in a minimally growing condition at 30°C by integrating the experimentally derived Hi-C interaction matrix within a computer model. Our quantitative analysis demonstrates that, while the dynamics of the chromosome is sub-diffusive in a viscoelastic media in general, the diffusion constants and the diffusive exponents are strongly dependent on the spatial coordinates of chromosomal loci. In particular, the loci in Ter Macro-domain display slower mobility compared to the others. The result is found to be robust even in the presence of active noise. Interestingly, a series of control investigations reveal that the absence of Hi-C interactions in the model would have abolished the heterogeneity in loci diffusion, indicating that the observed coordinate-dependent chromosome dynamics is heavily dictated via Hi-C-guided long-range inter-loci communications. Overall, the study underscores the key role of Hi-C interactions in guiding the inter-loci encounter and in modulating the underlying heterogeneity of the loci diffusion.

Structural basis of ALC1/CHD1L autoinhibition and the mechanism of activation by the nucleosome

Chromatin remodeler ALC1 (amplification in liver cancer 1) is crucial for repairing damaged DNA. It is autoinhibited and activated by nucleosomal epitopes. However, the mechanisms by which ALC1 is regulated remain unclear. Here we report the crystal structure of human ALC1 and the cryoEM structure bound to the nucleosome. The structure shows the macro domain of ALC1 binds to lobe 2 of the ATPase motor, sequestering two elements for nucleosome recognition, explaining the autoinhibition mechanism of the enzyme. The H4 tail competes with the macro domain for lobe 2-binding, explaining the requirement for this nucleosomal epitope for ALC1 activation. A dual-arginine-anchor motif of ALC1 recognizes the acidic pocket of the nucleosome, which is critical for chromatin remodeling in vitro. Together, our findings illustrate the structures of ALC1 and shed light on its regulation mechanisms, paving the way for the discovery of drugs targeting ALC1 for the treatment of cancer.

Structural basis of ALC1/CHD1L autoinhibition and the mechanism of activation by the nucleosome

Chromatin remodeler ALC1 (amplification in liver cancer 1) is crucial for repairing damaged DNA. It is autoinhibited and activated by nucleosomal epitopes. However, the mechanisms by which ALC1 is regulated remain unclear. Here we report the crystal structure of human ALC1 and the cryoEM structure bound to the nucleosome. The structure shows the macro domain of ALC1 binds to lobe 2 of the ATPase motor, sequestering two elements for nucleosome recognition, explaining the autoinhibition mechanism of the enzyme. The H4 tail competes with the macro domain for lobe 2-binding, explaining the requirement for this nucleosomal epitope for ALC1 activation. A dual-arginine-anchor motif of ALC1 recognizes the acidic pocket of the nucleosome, which is critical for chromatin remodeling in vitro. Together, our findings illustrate the structures of ALC1 and shed light on its regulation mechanisms, paving the way for the discovery of drugs targeting ALC1 for the treatment of cancer.

Conceptualizing change in organizational cognition

PurposeSecchi and Cowley (2016, 2018) propose a Radical approach to Organizational Cognition (ROC) as a way of studying cognitive processes in organizations. What distinguishes ROC from the established research on Organizational Cognition is that it remains faithful to radical, anti-representationalist principles of contemporary cognitive science. However, it is imperative for proponents of ROC to legitimize their approach by considering how it differs from the established research approach of Distributed Cognition (DCog). DCog is a potential contender to ROC in that it not only counters classical approaches to cognition but also provides valuable insights into cognition in organizational settings.Design/methodology/approachThe paper adopts a conceptual/theoretical approach that expands Secchi and Cowley's introduction of ROC.FindingsThe paper shows that DCog research presupposes a task-specification requirement, which entails that cognitive tasks are well-defined. Consequently, DCog research neglects cases of organizational becoming where tasks cannot be clearly demarcated for the or are well-known to the organization. This is the case with the introduction of novel tasks or technical devices. Moreover, the paper elaborates on ROC's 3M model by linking it with insights from the literature on organizational change. Thus, it explores how organizing can be explored as an emergent phenomenon that involves micro, meso and macro domain dynamics, which are shaped by synoptic and performative changes.Originality/valueThe present paper explores new grounds for ROC by not only expanding on its core model but also showing its potential for informing organizational theory and radical cognitive science research.

Elucidating the tunability of binding behavior for the MERS-CoV macro domain with NAD metabolites

The macro domain is an ADP-ribose (ADPR) binding module, which is considered to act as a sensor to recognize nicotinamide adenine dinucleotide (NAD) metabolites, including poly ADPR (PAR) and other small molecules. The recognition of macro domains with various ligands is important for a variety of biological functions involved in NAD metabolism, including DNA repair, chromatin remodeling, maintenance of genomic stability, and response to viral infection. Nevertheless, how the macro domain binds to moieties with such structural obstacles using a simple cleft remains a puzzle. We systematically investigated the Middle East respiratory syndrome-coronavirus (MERS-CoV) macro domain for its ligand selectivity and binding properties by structural and biophysical approaches. Of interest, NAD, which is considered not to interact with macro domains, was co-crystallized with the MERS-CoV macro domain. Further studies at physiological temperature revealed that NAD has similar binding ability with ADPR because of the accommodation of the thermal-tunable binding pocket. This study provides the biochemical and structural bases of the detailed ligand-binding mode of the MERS-CoV macro domain. In addition, our observation of enhanced binding affinity of the MERS-CoV macro domain to NAD at physiological temperature highlights the need for further study to reveal the biological functions.

Engineering Af1521 improves ADP-ribose binding and identification of ADP-ribosylated proteins

Protein ADP-ribosylation is a reversible post-translational modification that regulates important cellular functions. The identification of modified proteins has proven challenging and has mainly been achieved via enrichment methodologies. Random mutagenesis was used here to develop an engineered Af1521 ADP-ribose binding macro domain protein with 1000-fold increased affinity towards ADP-ribose. The crystal structure reveals that two point mutations K35E and Y145R form a salt bridge within the ADP-ribose binding domain. This forces the proximal ribose to rotate within the binding pocket and, as a consequence, improves engineered Af1521 ADPr-binding affinity. Its use in our proteomic ADP-ribosylome workflow increases the ADP-ribosylated protein identification rates and yields greater ADP-ribosylome coverage. Furthermore, generation of an engineered Af1521 Fc fusion protein confirms the improved detection of cellular ADP-ribosylation by immunoblot and immunofluorescence. Thus, this engineered isoform of Af1521 can also serve as a valuable tool for the analysis of cellular ADP-ribosylation under in vivo conditions.