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Structure and function of Semaphorin-5A glycosaminoglycan interactions

Abstract:
Integration of extracellular signals by neurons is pivotal for brain development, plasticity, and repair. Axon guidance relies on receptor-ligand interactions crosstalking with extracellular matrix components. Semaphorin-5A (Sema5A) is a bifunctional guidance cue exerting attractive and inhibitory effects on neuronal growth through the interaction with heparan sulfate (HS) and chondroitin sulfate (CS) glycosaminoglycans (GAGs), respectively. Sema5A harbors seven thrombospondin type-1 repeats (TSR1-7) important for GAG binding, however the underlying molecular basis and functions in vivo remain enigmatic. Here we dissect the structural basis for Sema5A:GAG specificity and demonstrate the functional significance of this interaction in vivo. Using x-ray crystallography, we reveal a dimeric fold variation for TSR4 that accommodates GAG interactions. TSR4 co-crystal structures identify binding residues validated by site-directed mutagenesis. In vitro and cell-based assays uncover specific GAG epitopes necessary for TSR association. We demonstrate that HS-GAG binding is preferred over CS-GAG and mediates Sema5A oligomerization. In vivo, Sema5A:GAG interactions are necessary for Sema5A function and regulate Plexin-A2 dependent dentate progenitor cell migration. Our study rationalizes Sema5A associated developmental and neurological disorders and provides mechanistic insights into how multifaceted guidance functions of a single transmembrane cue are regulated by proteoglycans.
Publication status:
Published
Peer review status:
Peer reviewed

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Institution:
University of Oxford
Role:
Author
ORCID:
0000-0003-1761-5610
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Role:
Author
ORCID:
0000-0003-0448-6135
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Role:
Author
ORCID:
0000-0002-3566-8698


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Funder identifier:
10.13039/501100009708
Grant:
NNF22OC0073736
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Funder identifier:
10.13039/501100002808
Grant:
CF20-0412
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Funder identifier:
10.13039/501100000265
Grant:
MR/T000503/1
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Funder identifier:
10.13039/100010269
Grant:
223133/Z/21/Z


Publisher:
Nature Research
Journal:
Nature Communications More from this journal
Volume:
15
Issue:
1
Pages:
2723-2723
Article number:
2723
Publication date:
2024-03-28
DOI:
EISSN:
2041-1723
ISSN:
2041-1723


Language:
English
Keywords:
Pubs id:
1924966
Local pid:
pubs:1924966
Source identifiers:
W4393258643
Deposit date:
2026-06-10
ARK identifier:
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