Cdc48 (p97 or VCP in metazoans) targets polyubiquitin to selectively disassemble and degrade proteins. Cdc48 is believed to move along the ubiquitin chain towards linked proteins, but this has not been directly observed. By following single molecules in time, we find that the polypeptide branch points of ubiquitin chains are repeatedly inserted and rejected from the Cdc48 pore in ATP-driven manner, in bursts lasting up to seconds. This non-processive mode either ends by terminal substrate rejection, or advances to processive action, which drives ubiquitin unfolding in two steps, branch point translocation, and the extrusion of ubiquitin and linked protein as polypeptide loops in trans. Final retrograde movement can bring polypeptide segments back to cis. Our results establish the dynamics of ubiquitin chain handling by Cdc48, and reveal key hallmarks of kinetic proofreading. We speculate that Cdc48 translocation may play a role in ubiquitin chain selection. ... Read more

Haptic teleoperation is a promising technology with applications in telemaintenance and disaster management. However, it faces significant challenges when the application is subjected to a high network latency and environments with moving objects. This work aims to extend Model Mediated Teleoperation (MMT) to overcome challenges in supporting dynamic environments. Instead of striving for perfect model alignment, we acknowledge the inevitable mismatch between the remote environment and its model at the operator. We propose a set of design principles and an accompanying framework for designing MMT solutions that prioritize operator intent. Our approach is exemplified through an application where an operator, located 8000 km away (The Netherlands - India) and subjected to an average of 179 ms end-to-end latency, guides a robot arm to draw on a whiteboard whose position is actively altered. We evaluate the effectiveness of our approach through a user study. We show a 3-point improvement on a 7-point Likert scale when users utilize our approach to teleoperate over significant network latency of up to 1 s. ... Read more

Delivery of naked DNA molecules into living cells via physical disruption of the membrane under electric pulses has potential biomedical applications ranging from gene electro-transfer, electro-chemotherapy, to gene therapy, yet the mechanisms involved in DNA transport remain vague. To investigate the mechanism of DNA translocation across the cell membrane, giant unilamellar vesicles (GUVs) were electroporated in the presence of DNA molecules keeping the size of the DNA molecules as a variable parameter. We experimentally determined the translocation efficiency for each size of the DNA molecule, to compare the results with the existing and conflicting theories of the translocation mechanism i.e. stochastic threading and bulk electrophoresis. We observed that the translocation efficiency is independent of DNA size (ranging from 25–20 000 bp, bp = base pairs), implying that DNA molecules translocate freely across the electro-pores in the lipid membrane in their native polymer conformation, as opposed to unravelling and threading through the electro-pore. Bulk electrophoretic mobility determines the relationship between translocation efficiency and the size of the DNA molecule. This research provides experimental evidence of the mechanistic understanding of DNA translocation across lipid membranes which is essential for devising efficient and predictable protocols for electric field mediated naked DNA delivery. ... Read more