Axon guidance relies on precise translation of extracellular signal gradients into local changes in cytoskeletal dynamics, but the molecular mechanisms regulating dose-dependent responses of growth cones are still poorly understood. Here, we show that during embryonic development in growing axons, a low level of Semaphorin3A stimulation is buffered by the prolyl isomerase Pin1. We demonstrate that Pin1 stabilizes CDK5-phosphorylated CRMP2A, the major isoform of CRMP2 in distal axons. Consequently, Pin1 knockdown or knockout reduces CRMP2A levels specifically in distal axons and inhibits axon growth, which can be fully rescued by Pin1 or CRMP2A expression. Moreover, Pin1 knockdown or knockout increases sensitivity to Sema3A-induced growth cone collapse in vitro and in vivo, leading to developmental abnormalities in axon guidance. These results identify an important isoform-specific function and regulation of CRMP2A in controlling axon growth and uncover Pin1-catalyzed prolyl isomerization as a regulatory mechanism in axon guidance.

Biomedical Research Institute University of Dundee Ninewells Hospital Dundee DD1 9SY Scotland UK

Department of Medicine Beth Israel Deaconess Medical Center Harvard Medical School 330 Brookline Avenue CLS 0408 Boston MA 02215 USA

Department of Medicine Beth Israel Deaconess Medical Center Harvard Medical School 330 Brookline Avenue CLS 0408 Boston MA 02215 USA; Institute for Translational Medicine Fujian Medical University Fuzhou 350108 China

Department of Medicine Beth Israel Deaconess Medical Center Harvard Medical School 330 Brookline Avenue CLS 0408 Boston MA 02215 USA; Institute of Molecular Genetics Vídeňská 1083 142 20 Prague 4 Czech Republic

Department of Medicine Beth Israel Deaconess Medical Center Harvard Medical School 330 Brookline Avenue CLS 0408 Boston MA 02215 USA; Institute of Molecular Genetics Vídeňská 1083 142 20 Prague 4 Czech Republic; Institute of Physiology Vídeňská 1083 142 20 Prague 4 Czech Republic

Department of Pediatric Oncology and Cancer Biology Dana Farber Cancer Institute Boston MA 02215 USA; Department of Neurobiology Harvard Medical School Boston MA 02115 USA

Institute of Molecular Genetics Vídeňská 1083 142 20 Prague 4 Czech Republic

Institute of Molecular Genetics Vídeňská 1083 142 20 Prague 4 Czech Republic; Institute of Physiology Vídeňská 1083 142 20 Prague 4 Czech Republic

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