Ubiquitin-protein ligase Ubr5 cooperates with hedgehog signalling to promote skeletal tissue homeostasis

David Mellis; Katherine A. Staines; Silvia Peluso; Ioanna Ch Georgiou; Natalie Dora; Malgorzata Kubiak; Rob van’t Hof; Michela Grillo; Colin Farquharson; Elaine Kinsella; Anna Thornburn; Stuart H. Ralston; Donald M. Salter; Natalia A. Riobo-Del Galdo; Robert E. Hill; Mark Ditzel

doi:10.1371/journal.pgen.1009275

Ubiquitin-protein ligase Ubr5 cooperates with hedgehog signalling to promote skeletal tissue homeostasis

David Mellis, Katherine A. Staines, Silvia Peluso, Ioanna Ch Georgiou, Natalie Dora, Malgorzata Kubiak, Rob van’t Hof, Michela Grillo, Colin Farquharson, Elaine Kinsella, Anna Thornburn, Stuart H. Ralston, Donald M. Salter, Natalia A. Riobo-Del Galdo, Robert E. Hill, Mark Ditzel

School of Applied Sciences

Research output: Contribution to journal › Article › peer-review

Abstract

Mammalian Hedgehog (HH) signalling pathway plays an essential role in tissue homeostasis and its deregulation is linked to rheumatological disorders. UBR5 is the mammalian homologue of the E3 ubiquitin-protein ligase Hyd, a negative regulator of the Hh-pathway in Drosophila. To investigate a possible role of UBR5 in regulation of the musculoskeletal system through modulation of mammalian HH signaling, we created a mouse model for specific loss of Ubr5 function in limb bud mesenchyme. Our findings revealed a role for UBR5 in maintaining cartilage homeostasis and suppressing metaplasia. Ubr5 loss of function resulted in progressive and dramatic articular cartilage degradation, enlarged, abnormally shaped sesamoid bones and extensive heterotopic tissue metaplasia linked to calcification of tendons and ossification of synovium. Genetic suppression of smoothened (Smo), a key mediator of HH signalling, dramatically enhanced the Ubr5 mutant phenotype. Analysis of HH signalling in both mouse and cell model systems revealed that loss of Ubr5 stimulated canonical HH-signalling while also increasing PKA activity. In addition, human osteoarthritic samples revealed similar correlations between UBR5 expression, canonical HH signalling and PKA activity markers. Our studies identified a crucial function for the Ubr5 gene in the maintenance of skeletal tissue homeostasis and an unexpected mode of regulation of the HH signalling pathway.

Original language	English
Article number	e1009275
Number of pages	25
Journal	PLoS Genetics
Volume	17
Issue number	4
DOIs	https://doi.org/10.1371/journal.pgen.1009275
Publication status	Published - 5 Apr 2021

Bibliographical note

Publisher Copyright:
© 2021 Mellis et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: MD is supported by a University of Edinburgh Chancellor’s Fellowship and funding from a Carnegie Research Incentive Grant (70356); BH by an MRC University Unit grant MM_UU_00007/8; NARD-G by the NIH (1R01GM088256) and a BBSRC project grant (BB/S01716X/1); KS by the MRC (MR/R022240/2) and CF by BBSRC through an Institute Strategic Programme Grant Funding (BB/J004316/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Keywords

Hedgehog signaling
Knee joints
chondrocytes
PKA signaling cascading
Cartilage
Signal processing
Tendons
Ossification

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Cite this

Mellis, D., Staines, K. A., Peluso, S., Georgiou, I. C., Dora, N., Kubiak, M., van’t Hof, R., Grillo, M., Farquharson, C., Kinsella, E., Thornburn, A., Ralston, S. H., Salter, D. M., Riobo-Del Galdo, N. A., Hill, R. E., & Ditzel, M. (2021). Ubiquitin-protein ligase Ubr5 cooperates with hedgehog signalling to promote skeletal tissue homeostasis. PLoS Genetics, 17(4), Article e1009275. https://doi.org/10.1371/journal.pgen.1009275

@article{f712c33762cc4c8ea67c347112b30a87,

title = "Ubiquitin-protein ligase Ubr5 cooperates with hedgehog signalling to promote skeletal tissue homeostasis",

abstract = "Mammalian Hedgehog (HH) signalling pathway plays an essential role in tissue homeostasis and its deregulation is linked to rheumatological disorders. UBR5 is the mammalian homologue of the E3 ubiquitin-protein ligase Hyd, a negative regulator of the Hh-pathway in Drosophila. To investigate a possible role of UBR5 in regulation of the musculoskeletal system through modulation of mammalian HH signaling, we created a mouse model for specific loss of Ubr5 function in limb bud mesenchyme. Our findings revealed a role for UBR5 in maintaining cartilage homeostasis and suppressing metaplasia. Ubr5 loss of function resulted in progressive and dramatic articular cartilage degradation, enlarged, abnormally shaped sesamoid bones and extensive heterotopic tissue metaplasia linked to calcification of tendons and ossification of synovium. Genetic suppression of smoothened (Smo), a key mediator of HH signalling, dramatically enhanced the Ubr5 mutant phenotype. Analysis of HH signalling in both mouse and cell model systems revealed that loss of Ubr5 stimulated canonical HH-signalling while also increasing PKA activity. In addition, human osteoarthritic samples revealed similar correlations between UBR5 expression, canonical HH signalling and PKA activity markers. Our studies identified a crucial function for the Ubr5 gene in the maintenance of skeletal tissue homeostasis and an unexpected mode of regulation of the HH signalling pathway.",

keywords = "Hedgehog signaling, Knee joints, chondrocytes, PKA signaling cascading, Cartilage, Signal processing, Tendons, Ossification",

author = "David Mellis and Staines, {Katherine A.} and Silvia Peluso and Georgiou, {Ioanna Ch} and Natalie Dora and Malgorzata Kubiak and {van{\textquoteright}t Hof}, Rob and Michela Grillo and Colin Farquharson and Elaine Kinsella and Anna Thornburn and Ralston, {Stuart H.} and Salter, {Donald M.} and {Riobo-Del Galdo}, {Natalia A.} and Hill, {Robert E.} and Mark Ditzel",

note = "Publisher Copyright: {\textcopyright} 2021 Mellis et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: MD is supported by a University of Edinburgh Chancellor{\textquoteright}s Fellowship and funding from a Carnegie Research Incentive Grant (70356); BH by an MRC University Unit grant MM_UU_00007/8; NARD-G by the NIH (1R01GM088256) and a BBSRC project grant (BB/S01716X/1); KS by the MRC (MR/R022240/2) and CF by BBSRC through an Institute Strategic Programme Grant Funding (BB/J004316/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.",

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doi = "10.1371/journal.pgen.1009275",

language = "English",

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Mellis, D, Staines, KA, Peluso, S, Georgiou, IC, Dora, N, Kubiak, M, van’t Hof, R, Grillo, M, Farquharson, C, Kinsella, E, Thornburn, A, Ralston, SH, Salter, DM, Riobo-Del Galdo, NA, Hill, RE & Ditzel, M 2021, 'Ubiquitin-protein ligase Ubr5 cooperates with hedgehog signalling to promote skeletal tissue homeostasis', PLoS Genetics, vol. 17, no. 4, e1009275. https://doi.org/10.1371/journal.pgen.1009275

TY - JOUR

T1 - Ubiquitin-protein ligase Ubr5 cooperates with hedgehog signalling to promote skeletal tissue homeostasis

AU - Mellis, David

AU - Staines, Katherine A.

AU - Peluso, Silvia

AU - Georgiou, Ioanna Ch

AU - Dora, Natalie

AU - Kubiak, Malgorzata

AU - van’t Hof, Rob

AU - Grillo, Michela

AU - Farquharson, Colin

AU - Kinsella, Elaine

AU - Thornburn, Anna

AU - Ralston, Stuart H.

AU - Salter, Donald M.

AU - Riobo-Del Galdo, Natalia A.

AU - Hill, Robert E.

AU - Ditzel, Mark

N1 - Publisher Copyright: © 2021 Mellis et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Funding: MD is supported by a University of Edinburgh Chancellor’s Fellowship and funding from a Carnegie Research Incentive Grant (70356); BH by an MRC University Unit grant MM_UU_00007/8; NARD-G by the NIH (1R01GM088256) and a BBSRC project grant (BB/S01716X/1); KS by the MRC (MR/R022240/2) and CF by BBSRC through an Institute Strategic Programme Grant Funding (BB/J004316/1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

PY - 2021/4/5

Y1 - 2021/4/5

N2 - Mammalian Hedgehog (HH) signalling pathway plays an essential role in tissue homeostasis and its deregulation is linked to rheumatological disorders. UBR5 is the mammalian homologue of the E3 ubiquitin-protein ligase Hyd, a negative regulator of the Hh-pathway in Drosophila. To investigate a possible role of UBR5 in regulation of the musculoskeletal system through modulation of mammalian HH signaling, we created a mouse model for specific loss of Ubr5 function in limb bud mesenchyme. Our findings revealed a role for UBR5 in maintaining cartilage homeostasis and suppressing metaplasia. Ubr5 loss of function resulted in progressive and dramatic articular cartilage degradation, enlarged, abnormally shaped sesamoid bones and extensive heterotopic tissue metaplasia linked to calcification of tendons and ossification of synovium. Genetic suppression of smoothened (Smo), a key mediator of HH signalling, dramatically enhanced the Ubr5 mutant phenotype. Analysis of HH signalling in both mouse and cell model systems revealed that loss of Ubr5 stimulated canonical HH-signalling while also increasing PKA activity. In addition, human osteoarthritic samples revealed similar correlations between UBR5 expression, canonical HH signalling and PKA activity markers. Our studies identified a crucial function for the Ubr5 gene in the maintenance of skeletal tissue homeostasis and an unexpected mode of regulation of the HH signalling pathway.

AB - Mammalian Hedgehog (HH) signalling pathway plays an essential role in tissue homeostasis and its deregulation is linked to rheumatological disorders. UBR5 is the mammalian homologue of the E3 ubiquitin-protein ligase Hyd, a negative regulator of the Hh-pathway in Drosophila. To investigate a possible role of UBR5 in regulation of the musculoskeletal system through modulation of mammalian HH signaling, we created a mouse model for specific loss of Ubr5 function in limb bud mesenchyme. Our findings revealed a role for UBR5 in maintaining cartilage homeostasis and suppressing metaplasia. Ubr5 loss of function resulted in progressive and dramatic articular cartilage degradation, enlarged, abnormally shaped sesamoid bones and extensive heterotopic tissue metaplasia linked to calcification of tendons and ossification of synovium. Genetic suppression of smoothened (Smo), a key mediator of HH signalling, dramatically enhanced the Ubr5 mutant phenotype. Analysis of HH signalling in both mouse and cell model systems revealed that loss of Ubr5 stimulated canonical HH-signalling while also increasing PKA activity. In addition, human osteoarthritic samples revealed similar correlations between UBR5 expression, canonical HH signalling and PKA activity markers. Our studies identified a crucial function for the Ubr5 gene in the maintenance of skeletal tissue homeostasis and an unexpected mode of regulation of the HH signalling pathway.

KW - Hedgehog signaling

KW - Knee joints

KW - chondrocytes

KW - PKA signaling cascading

KW - Cartilage

KW - Signal processing

KW - Tendons

KW - Ossification

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DO - 10.1371/journal.pgen.1009275

M3 - Article

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VL - 17

JO - PLoS Genetics

JF - PLoS Genetics

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ER -

Ubiquitin-protein ligase Ubr5 cooperates with hedgehog signalling to promote skeletal tissue homeostasis

Abstract

Bibliographical note

Keywords

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