Journal Article

KRAS interaction with RAF1 RAS-binding domain and cysteine-rich domain provides insights into RAS-mediated RAF activation

Young, Lucy C., Bindu, Lakshman, Nissley, Dwight V., Chan, Albert H., McCormick, Frank, Stephen, Andrew G., Taylor, Troy, Denson, John-Paul, Neale, Chris, Simanshu, Dhirendra K., Dharmaiah, Srisathiyanarayanan, Messing, Simon, Esposito, Dominic, Tran, Timothy H.

2021

Overview

The first step of RAF activation involves binding to active RAS, resulting in the recruitment of RAF to the plasma membrane. To understand the molecular details of RAS-RAF interaction, we present crystal structures of wild-type and oncogenic mutants of KRAS complexed with the RAS-binding domain (RBD) and the membrane-interacting cysteine-rich domain (CRD) from the N-terminal regulatory region of RAF1. Our structures reveal that RBD and CRD interact with each other to form one structural entity in which both RBD and CRD interact extensively with KRAS. Mutations at the KRAS-CRD interface result in a significant reduction in RAF1 activation despite only a modest decrease in binding affinity. Combining our structures and published data, we provide a model of RAS-RAF complexation at the membrane, and molecular insights into RAS-RAF interaction during the process of RAS-mediated RAF activation. The molecular details of the RAS-RAF interaction are still not fully understood. Here, the authors present crystal structures of wild-type and mutant KRAS in complex with the RAS-binding and membrane-interacting cysteine-rich domains of RAF1, and propose a model of the membrane-bound RAS-RAF complex.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 13/95 / 60 APPLIED LIFE SCIENCES / 631/45/612/1243 / 631/535/1266 / 631/67/395 / Binding / Binding Sites / Cancer / Crystal structure / Crystallography, X-Ray / Cysteine / Cysteine - metabolism / Domains / Humanities and Social Sciences / Humans / K-Ras protein / Kinases / Laboratories / Medical research / Membranes / Models, Molecular / multidisciplinary / Mutagenesis / Mutants / Mutation / Oncogene proteins / Oncogenes / Protein Binding / Protein Conformation / Protein Domains - genetics / Protein Interaction Domains and Motifs / Proteins / Proto-Oncogene Proteins c-raf - chemistry / Proto-Oncogene Proteins c-raf - metabolism / Proto-Oncogene Proteins p21(ras) - chemistry / Proto-Oncogene Proteins p21(ras) - genetics / Proto-Oncogene Proteins p21(ras) - metabolism / ras Proteins - chemistry / ras Proteins - metabolism / Science / Science (multidisciplinary) / X-ray crystallography