CENP-C unwraps the human CENP-A nucleosome through the H2A C-terminal tail

Centromeres are defined epigenetically by nucleosomes containing the histone H3 variant CENP-A, upon which the constitutive centromere-associated network of proteins (CCAN) is built. CENP-C is considered to be a central organizer of the CCAN. We provide new molecular insights into the structure of human CENP-A nucleosomes, in isolation and in complex with the CENP-C central region (CENP-C-CR), the main CENP-A binding module of human CENP-C. We establish that the short alpha N helix of CENP-A promotes DNA flexibility at the nucleosome ends, independently of the sequence it wraps. Furthermore, we show that, in vitro, two regions of human CENP-C (CENP-C-CR and CENP-C-motif) both bind exclusively to the CENP-A nucleosome. We find CENP-C-CR to bind with high affinity due to an extended hydrophobic area made up of CENP-A(V)(532) and CENP-A(V)(533). Importantly, we identify two key conformational changes within the CENP-A nucleosome upon CENP-C binding. First, the loose DNA wrapping of CENP-A nucleosomes is further exacerbated, through destabilization of the H2A C-terminal tail. Second, CENP-C-CR rigidifies the N-terminal tail of H4 in the conformation favoring H4(K20) monomethylation, essential for a functional centromere

Model for eukaryotic tail-anchored protein binding based on the structure of Get3

The Get3 ATPase directs the delivery of tail-anchored (TA) proteins to the endoplasmic reticulum (ER). TA-proteins are characterized by having a single transmembrane helix (TM) at their extreme C terminus and include many essential proteins, such as SNAREs, apoptosis factors, and protein translocation components. These proteins cannot follow the SRP-dependent co-translational pathway that typifies most integral membrane proteins; instead, post-translationally, these proteins are recognized and bound by Get3 then delivered to the ER in the ATP dependent Get pathway. To elucidate a molecular mechanism for TA protein binding by Get3 we have determined three crystal structures in apo and ADP forms from Saccharomyces cerevisae (ScGet3-apo) and Aspergillus fumigatus (AfGet3-apo and AfGet3-ADP). Using structural information, we generated mutants to confirm important interfaces and essential residues. These results point to a model of how Get3 couples ATP hydrolysis to the binding and release of TA-proteins

Drag reduction utilizing a wall-attached ferrofluid film in turbulent channel flow

This study explores the application of a wall-attached ferrofluid film to decrease skin friction drag in turbulent channel flow. We conduct experiments using water as a working fluid in a turbulent channel flow setup, where one wall is coated with a ferrofluid layer held in place by external permanent magnets. Depending on the flow conditions, the interface between the two fluids is observed to form unstable travelling waves. While ferrofluid coating has been previously employed in laminar and moderately turbulent flows to reduce drag by creating a slip condition at the fluid interface, its effectiveness in fully developed turbulent conditions, particularly when the interface exhibits instability, remains uncertain. Our primary objective is to assess the effectiveness of ferrofluid coating in reducing turbulent drag with particular focus on scenarios when the ferrofluid layer forms unstable waves. To achieve this, we measure flow velocity using two-dimensional particle tracking velocimetry (2D-PTV), and the interface contour between the fluids is determined using an interface tracking algorithm. Our results reveal the significant potential of ferrofluid coating for drag reduction, even in scenarios where the interface between the surrounding fluid and ferrofluid exhibits instability. In particular, waves with an amplitude significantly smaller than a viscous length scale positively contribute to drag reduction, while larger waves are detrimental, because of induced turbulent fluctuations. However, for the latter case, slip out-competes the extra turbulence so that drag is still reduced

Lagrangian coherent structures and entrainment near the turbulent/non-Turbulent interface of a gravity current

ISSN:0022-1120ISSN:1469-764

Universal alignment in turbulent pair dispersion

Countless processes in nature and industry, from rain droplet nucleation to plankton interaction in the ocean, are intimately related to turbulent fluctuations of local concentrations of advected matter. These fluctuations can be described by considering the change of the separation between particle pairs, known as pair dispersion, which is believed to obey a cubic in time growth according to Richardson's theory. Our work reveals a universal, scale-invariant alignment between the relative velocity and position vectors of dispersing particles at a mean angle that we show to be a universal constant of turbulence. We connect the value of this mean angle to Richardson's traditional theory and find agreement with data from a numerical simulation and a laboratory experiment. While the Richardson's cubic regime has been observed for small initial particle separations only, the constancy of the mean angle manifests throughout the entire inertial range of turbulence. Thus, our work reveals the universal nature of turbulent pair dispersion through a geometrical paradigm whose validity goes beyond the classical theory, and provides a novel framework for understanding and modeling transport and mixing processes.Comment: 20 pages, three figure

Co- and post-translational translocation through the protein-conducting channel:analogous mechanisms at work?

Many proteins are translocated across, or integrated into, membranes. Both functions are fulfilled by the 'translocon/translocase', which contains a membrane-embedded proteinconducting channel (PCC) and associated soluble factors that drive translocation and insertion reactions using nucleotide triphosphates as fuel. This perspective focuses on reinterpreting existing experimental data in light of a recently proposed PCC model comprising a front-to-front dimer of SecY or Sec61 heterotrimeric complexes. In this new framework, we propose (i) a revised model for SRP-SR-mediated docking of the ribosome-nascent polypeptide to the PCC; (ii) that the dynamic interplay between protein substrate, soluble factors and PCC controls the opening and closing of a transmembrane channel across, and/or a lateral gate into, the membrane; and (iii) that co-and post-translational translocation, involving the ribosome and SecA, respectively, not only converge at the PCC but also use analogous mechanisms for coordinating protein translocation

Structural basis of signal sequence surveillance and selection by the SRP–FtsY complex

Signal-recognition particle (SRP)-dependent targeting of translating ribosomes to membranes is a multistep quality-control process. Ribosomes that are translating weakly hydrophobic signal sequences can be rejected from the targeting reaction even after they are bound to the SRP. Here we show that the early complex, formed by Escherichia coli SRP and its receptor FtsY with ribosomes translating the incorrect cargo EspP, is unstable and rearranges inefficiently into subsequent conformational states, such that FtsY dissociation is favored over successful targeting. The N-terminal extension of EspP is responsible for these defects in the early targeting complex. The cryo-electron microscopy structure of this 'false' early complex with EspP revealed an ordered M domain of SRP protein Ffh making two ribosomal contacts, and the NG domains of Ffh and FtsY forming a distorted, flexible heterodimer. Our results provide a structural basis for SRP-mediated signal-sequence selection during recruitment of the SRP receptor