Multi-modal adaptor-clathrin contacts drive coated vesicle assembly.

Clathrin-coated pits are formed by the recognition of membrane and cargo by the AP2 complex and the subsequent recruitment of clathrin triskelia. A role for AP2 in coated-pit assembly beyond initial clathrin recruitment has not been explored. Clathrin binds the ß2 subunit of AP2, and several binding sites have been identified, but our structural knowledge of these interactions is incomplete and their functional importance during endocytosis is unclear. Here, we analysed the cryo-EM structure of clathrin cages assembled in the presence of ß2 hinge-appendage (ß2HA). We find that the ß2-appendage binds in at least two positions in the cage, demonstrating that multi-modal binding is a fundamental property of clathrin-AP2 interactions. In one position, ß2-appendage cross-links two adjacent terminal domains from different triskelia. Functional analysis of ß2HA-clathrin interactions reveals that endocytosis requires two clathrin interaction sites: a clathrin-box motif on the hinge and the "sandwich site" on the appendage. We propose that ß2-appendage binding to more than one triskelion is a key feature of the system and likely explains why assembly is driven by AP2.

Clathrin: the molecular shape shifter.

Clathrin is best known for its contribution to clathrin-mediated endocytosis yet it also participates to a diverse range of cellular functions. Key to this is clathrin's ability to assemble into polyhedral lattices that include curved football or basket shapes, flat lattices or even tubular structures. In this review, we discuss clathrin structure and coated vesicle formation, how clathrin is utilised within different cellular processes including synaptic vesicle recycling, hormone desensitisation, spermiogenesis, cell migration and mitosis, and how clathrin's remarkable 'shapeshifting' ability to form diverse lattice structures might contribute to its multiple cellular functions.

On-road and simulated driving: concurrent and discriminant validation.

INTRODUCTION: A converging pair of studies investigated the validity of a simulator for measuring driving performance/skill. STUDY 1: A concurrent validity study compared novice driver performance during an on-road driving test with their performance on a comparable simulated driving test. RESULTS: Results showed a reasonable degree of concordance in terms of the distribution of driving errors on-road and errors on the simulator. Moreover, there was a significant relationship between the two when driver performance was rank ordered according to errors, further establishing the relative validity of the simulator. However, specific driving errors on the two tasks were not closely related suggesting that absolute validity could not be established and that overall performance is needed to establish the level of skill. STUDY 2: A discriminant validity study compared driving performance on the simulator across three groups of drivers who differ in their level of experience--a group of true beginners who had no driving experience, a group of novice drivers who had completed driver education and had a learner's permit, and a group of fully licensed, experienced drivers. RESULTS: The findings showed significant differences among the groups in the expected direction--the various measures of driving errors showed that beginners performed worse than novice drivers and that experienced drivers had the fewest errors. Collectively, the results of the concurrent and discriminant validity studies support the use of the simulator as a valid measure of driving performance for research purposes. IMPACT ON INDUSTRY: These findings support the use of a driving simulator as a valid measure of driving performance for research purposes. Future research should continue to examine validity between on-road driving performance and performance on a driving simulator and the use of simulated driving tests in the evaluation of driver education/training programs.