The stability of R-spine defines RAF inhibitor resistance: A comprehensive analysis of oncogenic BRAF mutants with in-frame insertion of αC-ß4 loop.

Although targeting BRAF mutants with RAF inhibitors has achieved promising outcomes in cancer therapy, drug resistance remains a remarkable challenge, and underlying molecular mechanisms are not fully understood. Here, we characterized a previously unknown group of oncogenic BRAF mutants with in-frame insertions (LLRins506 or VLRins506) of αC-ß4 loop. Using structure modeling and molecular dynamics simulation, we found that these insertions formed a large hydrophobic network that stabilizes R-spine and thus triggers the catalytic activity of BRAF. Furthermore, these insertions disrupted BRAF dimer interface and impaired dimerization. Unlike BRAF(V600E), these BRAF mutants with low dimer affinity were strongly resistant to all RAF inhibitors in clinic or clinical trials, which arises from their stabilized R-spines. As predicted by molecular docking, the stabilized R-spines in other BRAF mutants also conferred drug resistance. Together, our data indicated that the stability of R-spine but not dimer affinity determines the RAF inhibitor resistance of oncogenic BRAF mutants.

Looking at remembering: Eye movements, pupil size, and autobiographical memory.

To examine the relationship between visual imagery and autobiographical memory, eye position and pupil size were recorded while participants first searched for memories and then reconstructed the retrieved memories (Experiment 1), or only searched for memories (Experiment 2). In Experiment 1, we observed that, although recollective experience was not associated with the number of fixations per minute, memories that took longer to retrieve were linked to increased pupil size. In Experiment 2, we observed that directly retrieved memories were recalled more quickly and were accompanied by smaller pupils than generatively retrieved memories. After correcting for response time, retrieval mode also produced an effect, showing that decreased pupil size is not simply due to directly retrieved memories being recalled more quickly. These findings provide compelling evidence that objective measures, such as pupil size, can be used alongside subjective measures, such as self-reports, to distinguish between directly retrieved and generatively retrieved memories.