Sonic Hedgehog Shedding Results in Functional Activation of the Solubilized Protein

Ohlig, Stefanie; Farshi, Pershang; Pickhinke, Ute; Boom, Johannes van den; Höing, Susanne; Jakuschev, Stanislav; Hoffmann, Daniel; Dreier, Rita; Schöler, Hans R.; Dierker, Tabea; Bordych, Christian; Grobe, Kay

doi:10.1016/j.devcel.2011.05.010

Cited by 83 publications

(131 citation statements)

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“…Despite small differences in the relative abundance of lower and higher molecular weight fractions, all ShhNp mutants multimerized effectively. We thus concluded that all ShhNp mutant forms assemble properly on the cell surface and are exclusively released in multimeric form (29).…”

Section: Shhn and Shhnp Binding To Heparin Ismentioning

confidence: 99%

Dual Roles of the Cardin-Weintraub Motif in Multimeric Sonic Hedgehog

Farshi¹,

Ohlig²,

Pickhinke³

et al. 2011

Journal of Biological Chemistry

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The fly morphogen Hedgehog (Hh) and its mammalian orthologs, Sonic, Indian, and Desert hedgehog, are secreted signaling molecules that mediate tissue patterning during embryogenesis and function in tissue homeostasis and regeneration in the adult. The function of all Hh family members is regulated at the levels of morphogen multimerization on the surface of producing cells, multimer release, multimer diffusion to target cells, and signal reception. These mechanisms are all known to depend on interactions of positively charged Hh amino acids (the Cardin-Weintraub (CW) motif) with negatively charged heparan sulfate (HS) glycosaminoglycan chains. However, a precise mechanistic understanding of these interactions is still lacking. The proteins of the Hh family are powerful morphogens that control growth and patterning of developing embryos. Current models for Hh activity suggest that the morphogen disperses from a localized source and forms a gradient that patterns fields of responsive cells expressing the Hh receptor Patched (Ptc).Genetic evidence suggests that this process critically depends on heparan sulfate proteoglycan (HSPG) 2 expression. Upon secretion to the cell surface, Drosophila Hh forms nanoscale oligomers on the cell surface that co-localize with HSPGs (1). HS binds to the Cardin-Weintraub (CW) motif found on all known Hhs and regulates their function in flies (2, 3) and mice (4). In Drosophila (5) as well as in mammalian cell culture (6, 7), Hhs are always released from producing cells in multimeric form, as demonstrated by gel filtration analysis of the soluble morphogen. Release of the multimeric morphogen (the processed Hh N-terminal signaling domain, designated HhNp) from the cell surface depends on the expression of Dispatched (8) and A disintegrin and metalloprotease (ADAM) family members that mediate ectodomain shedding from transfected Bosc23 cells (9). HS is involved in the formation of the HhNp extracellular gradient, which, in the fly, depends on the expression of the Drosophila Exostosin (Ext) family of proteins, encoded by the genes tout velu (ttv), brother of tout velu, and sister of tout velu and the glycosylphosphatidylinositollinked HSPGs Dally and Dally-like, corresponding to vertebrate glypicans (2, 3, 10). HS expression and Dally-like/glypican expression are also essential for signal reception and modulation on Ptc-expressing receiving cells (10 -14) and participate in HhNp-Ihog interaction (15). However, the essential role of direct morphogen-HSPG interactions in embryonic patterning was recently challenged (16). In that report, transgenic mice made deficient in two ShhNp CW amino acid residues implicated in HS binding (17) lacked an Shh-related phenotype, suggesting that direct morphogen-HS interactions were not essential for normal development. However, in that report as well as in others (16 -18), CW-dependent HS interactions were characterized using a recombinant, non-physiological monomeric morphogen termed ShhN, whereas embryogenesis depends entirely on the activity of morp...

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Section: Shhn and Shhnp Binding To Heparin Ismentioning

confidence: 99%

Dual Roles of the Cardin-Weintraub Motif in Multimeric Sonic Hedgehog

Farshi¹,

Ohlig²,

Pickhinke³

et al. 2011

Journal of Biological Chemistry

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“…2B, asterisk). These results demonstrate that both Hh lipidations contribute to Hh association with the cell membrane (Chamoun et al, 2001;Konitsiotis et al, 2014;Ohlig et al, 2011) to ensure that only fully processed Hh is released (Ohlig et al, 2012). This raises the question of how Hh shedding is controlled.…”

Section: Loss Of Dally Function In Hh-producing Cells Affects Drosophmentioning

confidence: 87%

“…2A). We and others have shown that proteolytic removal of N-and C-terminal lipidated peptides (called shedding) releases bioactive Shh morphogens from the surface of producing cells in vitro (Damhofer et al, 2015;Dierker et al, 2009;Jakobs et al, 2014;Ohlig et al, 2011). We further showed that because of the lack of the N-terminal Hhat acceptor cysteine (C25S or C25A in mouse Shh) (Hardy and Resh, 2012) non-palmitoylated Hh proteins require only C-terminal shedding for their solubilization, whereas most Shh N-termini are left intact (Jakobs et al, 2014).…”

Section: Loss Of Dally Function In Hh-producing Cells Affects Drosophmentioning

confidence: 95%

“…One group of molecules involved in Shh solubilization are a disintegrin and metalloproteinase (ADAM) sheddases (Dierker et al, 2009;Ohlig et al, 2011). Sheddases release numerous membrane-associated molecules from the cell surface (summarized in Tellier et al, 2006;Yang et al, 2006), while the cell retains the membrane-associated remnant peptides.…”

Section: Introductionmentioning

confidence: 99%

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Sonic hedgehog processing and release are regulated by glypican heparan sulfate proteoglycans

Ortmann,

Pickhinke,

Exner

et al. 2015

Journal of Cell Science

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There was an error published in J. Cell Sci. 128, 2374-2385.The reference Oustah et al. (2014) was cited incorrectly throughout the manuscript and in the reference list. Citations of Oustah et al. (2014) should read Al Oustah et al. (2014), and the correct reference is given below.Al Oustah, A., Danesin, C., Khouri-Farah, N., Farreny, M.-A., Escalas, N., Cochard, P., Glise, B. and Soula, C. (2014 show that glypican heparan sulfate proteoglycans regulate this process, through their heparan sulfate chains, in a cell autonomous manner. Heparan sulfate specifically modifies Shh processing at the cell surface, and purified glycosaminoglycans enhance the proteolytic removal of N-and C-terminal Shh peptides under cellfree conditions. The most likely explanation for these observations is direct Shh processing in the extracellular compartment, suggesting that heparan sulfate acts as a scaffold or activator for Shh ligands and the factors required for their turnover. We also show that purified heparan sulfate isolated from specific cell types and tissues mediates the release of bioactive Shh from pancreatic cancer cells, revealing a previously unknown regulatory role for these versatile molecules in a pathological context.

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“…Urinary concentrations of ADAM12 have been shown to correlate with cancer stage, making it a potential biomarker with which to monitor tumor progression (59)(60)(61). ADAM12 facilitates tumor progression by stimulating cell proliferation and survival pathways (62)(63)(64). This is achieved by the proteolytic activation and release (ectodomain shedding) of membrane-bound growth factors, including HB-EGF, erythroblastic leukemia viral oncogene homolg 4, tumor necrosis factor α and tumor growth factor α (40,65).…”

Section: Discussionmentioning

confidence: 99%

Effects of nitric oxide synthase deficiency on a disintegrin and metalloproteinase domain-containing protein 12 expression in mouse brain samples

Lendeckel¹,

Wolke²,

Bernstein

et al. 2015

Molecular Medicine Reports

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Abstract.A d isi nt eg r i n a nd met a l loprot ei nase domain-containing protein 12 (ADAM12) belongs to the ADAM family of transmembrane proteins. Via proteolysis, cell adhesion, cell-cell fusion, cell-matrix interaction and membrane protein shedding, ADAM proteins are involved in normal brain development, and also in cancer genesis and progression, and in inflammation. Therefore, neurobiological research focusing on this protein is increasing. Nitric oxide (NO), which is endogenously produced by NO synthases (NOS), is associated with glial tumors. However, knock-out of NOS produces only limited antitumor effects. The present study analyzed the expression of ADAM12 in the cortex and hippocampus of C57/BL6 wild-type mice, and endothelial NOS-, neuronal NOS-(nNOS) or inducible NOS (iNOS)-deficient ( -/-) mice, at different stages of development. Expression of ADAM12 was quantified using immunoblot analysis of cortical and hippocampal tissue samples from fetal, neonatal (5 days postnatal), adult (12 weeks old) or >1 year old mice. Using reverse transcription-quantitative polymerase chain reaction, ADAM12 expression was analyzed in cultured N9, OLN93, C6 and PC12 cells, representing the four main cell types in the brain, following NOS inhibition. ADAM12 expression was low in all mouse genotypes and regions of the brain, and in fetal and neonatal mice, an increase in expression was observed with increasing age. The highest levels of expression were observed in the cortex of adult mice, iNOS -/-mice of >1 year and wild-type mice, and in the hippocampus of adult and iNOS -/-mice of >1 year. By contrast, ADAM12 expression was lowest in adult nNOS -/-mice. Inhibition of NOS using N ω -Nitro-L-arginine methyl ester hydrochloride, induced ADAM12 mRNA expression in N9 and PC12 cell lines. Inhibition of NOS using L-N 6 -(1-Iminoethyl)lysine dihydrochloride, induced ADAM12 mRNA expression in N9 and C6 cell lines. No change in ADAM12 expression was observed in OLN93 cells following NOS inhibition. ADAM12 expression in mouse hippocampus and cortex samples demonstrated considerable variation during development, with a marked increase observed in adult and >1 year old mice, compared with that in fetal and neonatal mice. IntroductionNitric oxide (NO) is an important signaling molecule found in animals, including humans. Reduced NO production is associated with important cardiovascular risk factors, such as hyperlipidemia (1,2), diabetes, hypertension, smoking, atherosclerosis and aging (3). The bioavailability of NO in cells and tissues decreases with age. This may be a result of a decrease in the expression of the constitutive isoforms [endothelial nitric oxide synthase (eNOS) and neuronal NOS (nNOS)], with age (4). Inducible NOS (iNOS)-mediated NO formation has been shown to affect longevity. In mice, iNOS overexpression may lead to increased mortality, which is associated with cardiac hypertrophy and sudden cardiac death as a result of bradyarrhythmias (5). iNOS expression is typically not observed in the brains of young (1-3...

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Sonic Hedgehog Shedding Results in Functional Activation of the Solubilized Protein

Cited by 83 publications

References 50 publications

Dual Roles of the Cardin-Weintraub Motif in Multimeric Sonic Hedgehog

Dual Roles of the Cardin-Weintraub Motif in Multimeric Sonic Hedgehog

Sonic hedgehog processing and release are regulated by glypican heparan sulfate proteoglycans

Effects of nitric oxide synthase deficiency on a disintegrin and metalloproteinase domain-containing protein 12 expression in mouse brain samples

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