Secreted HHIP1 interacts with heparan sulfate and regulates Hedgehog ligand localization and function

Holtz, Alexander M.; Griffiths, Samuel C.; Davis, Samantha; Bishop, B.; Siebold, Christian; Allen, Benjamin L.

doi:10.1083/jcb.201411024

Cited by 40 publications

(40 citation statements)

References 113 publications

(152 reference statements)

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“…2E) (Chuang and McMahon, 1999). HHIP associates with the cell surface and extracellular matrix by binding to HSPGs (Holtz et al, 2015). An interesting question is why are CDO and BOC ligand agonists whereas HHIP is a ligand antagonist, even though both proteins bind to the same surface of SHH?…”

Section: The Interaction Of Hh Ligands With Co-receptors and Antagonistsmentioning

confidence: 99%

Biochemical mechanisms of vertebrate hedgehog signaling

2019

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Signaling pathways that mediate cell-cell communication are essential for collective cell behaviors in multicellular systems. The hedgehog (HH) pathway, first discovered and elucidated in Drosophila, is one of these iconic signaling systems that plays many roles during embryogenesis and in adults; abnormal HH signaling can lead to birth defects and cancer. We review recent structural and biochemical studies that have advanced our understanding of the vertebrate HH pathway, focusing on the mechanisms by which the HH signal is received by patched on target cells, transduced across the cell membrane by smoothened, and transmitted to the nucleus by GLI proteins to influence gene-expression programs.

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Section: The Interaction Of Hh Ligands With Co-receptors and Antagonistsmentioning

confidence: 99%

Biochemical mechanisms of vertebrate hedgehog signaling

2019

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“…Alternatively, they might result from Hhip1 moving towards the NT. Indeed, in spite of the initial findings that Hhip1 is a transmembrane glycoprotein with cell autonomous functions [18,30], it has recently been reported that Hhip1 is a secreted Shh antagonist able to exert long range effects on Shh signaling [31,32]. This is compatible with the possibility that sclerotomal Hhip1 crosses into the NT or at least localizes to its surrounding basement membrane [31].…”

Section: Reduction Of Shh In Sclerotome By Hhip1 Affects Both Myotomementioning

confidence: 98%

“…To discriminate between the above possibilities, we produced a Hhip:CD4 plasmid encoding for a membrane-tethered version of Hhip1, unable to undergo secretion [31,32]. First, we asked whether NT or sclerotomal cells missexpressing either Hhip1 or Hhip:CD4 are able to sequester Shh.…”

Section: Vis-à-vis Shhmentioning

confidence: 99%

“…Expression vectors were: pCAGGS-GFP, pCAGGS-RFP [77], Ptc1 [78], PTC Δloop2 [23,78], a retinoic acid reporter fused to alkaline phosphatase (pRARE-AP, from J. Sen) [79], a dominant negative pan-retinoic acid receptor (RAR403dn-IRES-GFP, from S. Sockanathan) that abrogates retinoic acid signaling [80], full length Shh [23], cholesterol deficient Shh (mShh-N-YFP, from V. Wallace) [33] that was subcloned into pCAGGS, and Hhip1 [23]. To produce Hhip1:CD4, a membrane-tethered version of Hhip1, the transmembrane and intracellular domains of mouse CD4 were fused to the C-terminal domain of Hhip1 lacking amino acids A679-V700, as previously described [31,32] and further subcloned into pCAGGS for electroporation. Electroporations were performed to the ventral half of epithelial somites (prospective sclerotome).…”

Section: Expression Vectors and Electroporationmentioning

confidence: 99%

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Neural tube development depends on notochord-derived Sonic hedgehog released into the sclerotome

Kahane

Kalcheim²

2019

Preprint

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AbstractSonic hedgehog (Shh), produced in notochord and floor plate, is necessary both for neural and mesodermal development. To reach the myotome, Shh has to traverse the sclerotome. By loss and gain of Shh function, and floor plate deletions, we report that sclerotomal Shh is also necessary for neural tube development. Reducing the amount of Shh in sclerotome by membrane-tethered hedgehog-interacting protein or by Patched1, but not by dominant active Patched, decreased motoneuron numbers while also compromising myotome differentiation. These effects were a specific and direct consequence of reducing Shh. In addition, grafting notochords in a basal, but not apical location vis-a-vis the tube, profoundly affected motoneuron development, suggesting that initial ligand presentation occurs at the basal side of epithelia corresponding to the sclerotome-neural tube interface.Collectively, our results reveal that the sclerotome is a potential site of a Shh gradient that coordinates development of mesodermal and neural progenitors.Summary statementShh that transits through the sclerotome is presented to the neuroepithelium from its basal aspect to affect motoneuron development.

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“…Using these criteria, a total of 32 cell-surface or extracellular proteins were confidently identified in five independent experiments using proteinase K or chymotrypsin (Table S1). They included known HSBPs, such as thrombospondin 1 (THBS1) (16 -18), hedgehog-interacting protein (HHIP) (19), and vascular endothelial growth factor receptor 1 (VEGFR1) (20). Previously unknown HSBPs were also detected, including C-type lectin domain family 14 member A (CLEC14A), tyrosine receptor phosphatase ␤ (PTPR␤), lysyl oxidase-like protein 2 (LOXL2), transmembrane protein 132 (TMEM132A), growth/ differentiation factor 15 (GDF15), adhesion G-proteincoupled receptor L2 (ADGRL2), and killer cell immunoglobulin-like receptor 3DL2 (KIR3DLR2).…”

Section: Analysis Of Heparin-binding Proteins On the Surface Of Endotmentioning

confidence: 99%

Proteomics-based screening of the endothelial heparan sulfate interactome reveals that C-type lectin 14a (CLEC14A) is a heparin-binding protein

Sandoval

Toledo

Painter

et al. 2020

Journal of Biological Chemistry

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Animal cells express heparan sulfate proteoglycans that perform many important cellular functions by way of heparan sulfate–protein interactions. The identification of membrane heparan sulfate–binding proteins is challenging because of their low abundance and the need for extensive enrichment. Here, we report a proteomics workflow for the identification and characterization of membrane-anchored and extracellular proteins that bind heparan sulfate. The technique is based on limited proteolysis of live cells in the absence of denaturation and fixation, heparin-affinity chromatography, and high-resolution LC-MS/MS, and we designate it LPHAMS. Application of LPHAMS to U937 monocytic and primary murine and human endothelial cells identified 55 plasma membrane, extracellular matrix, and soluble secreted proteins, including many previously unidentified heparin-binding proteins. The method also facilitated the mapping of the heparin-binding domains, making it possible to predict the location of the heparin-binding site. To validate the discovery feature of LPHAMS, we characterized one of the newly-discovered heparin-binding proteins, C-type lectin 14a (CLEC14A), a member of the C-type lectin family that modulates angiogenesis. We found that the C-type lectin domain of CLEC14A binds one-to-one to heparin with nanomolar affinity, and using molecular modeling and mutagenesis, we mapped its heparin-binding site. CLEC14A physically interacted with other glycosaminoglycans, including endothelial heparan sulfate and chondroitin sulfate E, but not with neutral or sialylated oligosaccharides. The LPHAMS technique should be applicable to other cells and glycans and provides a way to expand the repertoire of glycan-binding proteins for further study.

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Secreted HHIP1 interacts with heparan sulfate and regulates Hedgehog ligand localization and function

Abstract: HHIP1 acts as a secreted Hedgehog antagonist whose association with heparan sulfate–containing basement membranes regulates Hedgehog ligand distribution.

Cited by 40 publications

References 113 publications

Biochemical mechanisms of vertebrate hedgehog signaling

Biochemical mechanisms of vertebrate hedgehog signaling

Neural tube development depends on notochord-derived Sonic hedgehog released into the sclerotome

Proteomics-based screening of the endothelial heparan sulfate interactome reveals that C-type lectin 14a (CLEC14A) is a heparin-binding protein

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