Sensory deficit screen identifies nsf mutation that differentially affects SNARE recycling and quality control

Yan Gao; Yousuf A Khan; Weike Mo; K Ian White; Matthew Perkins; Richard A Pfuetzner; Josef G Trapani; Axel T Brunger; Teresa Nicolson

doi:10.1016/j.celrep.2023.112345

Sensory deficit screen identifies nsf mutation that differentially affects SNARE recycling and quality control

Cell Rep. 2023 Apr 25;42(4):112345. doi: 10.1016/j.celrep.2023.112345. Epub 2023 Apr 5.

Authors

Yan Gao¹, Yousuf A Khan², Weike Mo³, K Ian White⁴, Matthew Perkins⁵, Richard A Pfuetzner⁴, Josef G Trapani⁵, Axel T Brunger⁴, Teresa Nicolson⁶

Affiliations

¹ Department of Otolaryngology, Head and Neck Surgery, Stanford Medical School, 300 Pasteur Drive, Stanford, CA 94303, USA.
² Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA, USA; Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA, USA; Department of Structural Biology, Stanford University, Stanford, CA, USA; Department of Photon Science, Stanford University, Stanford, CA, USA; Center for Biomedical Informatics Research, Stanford University, Stanford, CA, USA.
³ Graduate Program Biomedical Sciences, Oregon Hearing Research Center and Vollum Institute, Oregon Health and Science University, 3181 SW Sam Jackson Park Road, Portland, OR 97239, USA.
⁴ Department of Molecular and Cellular Physiology, Stanford University, Stanford, CA, USA; Department of Neurology and Neurological Sciences, Stanford University, Stanford, CA, USA; Department of Structural Biology, Stanford University, Stanford, CA, USA; Department of Photon Science, Stanford University, Stanford, CA, USA; Howard Hughes Medical Institute, Stanford University, Stanford, CA, USA.
⁵ Department of Biology and Neuroscience Program, Amherst College, Amherst, MA 01002, USA.
⁶ Department of Otolaryngology, Head and Neck Surgery, Stanford Medical School, 300 Pasteur Drive, Stanford, CA 94303, USA. Electronic address: tnicolso@stanford.edu.

Abstract

The AAA⁺ NSF complex is responsible for SNARE complex disassembly both before and after membrane fusion. Loss of NSF function results in pronounced developmental and degenerative defects. In a genetic screen for sensory deficits in zebrafish, we identified a mutation in nsf, I209N, that impairs hearing and balance in a dosage-dependent manner without accompanying defects in motility, myelination, and innervation. In vitro experiments demonstrate that while the I209N NSF protein recognizes SNARE complexes, the effects on disassembly are dependent upon the type of SNARE complex and I209N concentration. Higher levels of I209N protein produce a modest decrease in binary (syntaxin-SNAP-25) SNARE complex disassembly and residual ternary (syntaxin-1A-SNAP-25-synaptobrevin-2) disassembly, whereas at lower concentrations binary disassembly activity is strongly reduced and ternary disassembly activity is absent. Our study suggests that the differential effect on disassembly of SNARE complexes leads to selective effects on NSF-mediated membrane trafficking and auditory/vestibular function.

Keywords: CP: Neuroscience; NSF; SNARE complex; balance; deafness; hair cell; hearing; ribbon synapse; sensory deficit; zebrafish.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Membrane Fusion*
Mutation / genetics
N-Ethylmaleimide-Sensitive Proteins / metabolism
Quality Control
SNARE Proteins* / genetics
SNARE Proteins* / metabolism
Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / genetics
Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / metabolism
Zebrafish / genetics
Zebrafish / metabolism

Substances

SNARE Proteins
Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
N-Ethylmaleimide-Sensitive Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding