Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
The deubiquitinating enzymes USP4 and USP17 target hyaluronan synthase 2 and differentially affect its function
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Ludwig Institute for Cancer Research. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Ludwig Institute for Cancer Research. Uppsala University, Science for Life Laboratory, SciLifeLab. AC Camargo Canc Ctr, Int Res Ctr, Dept Genom & Mol Biol, Sao Paulo, Brazil.; Univ Sao Paulo, Fac Med, Dept Pathol, Sao Paulo, Brazil.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Ludwig Institute for Cancer Research. Uppsala University, Science for Life Laboratory, SciLifeLab.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, Ludwig Institute for Cancer Research. Uppsala University, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0002-9508-896X
Show others and affiliations
2017 (English)In: Oncogenesis, E-ISSN 2157-9024, Vol. 6, article id e348Article in journal (Refereed) Published
Abstract [en]

The levels of hyaluronan, a ubiquitous glycosaminoglycan prominent in the extracellular matrix, is balanced through the actions of hyaluronan-synthesizing enzymes (HAS1, 2 and 3) and degrading hyaluronidases (Hyal 1, 2, 3 and PH20). Hyaluronan accumulates in rapidly remodeling tissues, such as breast cancer, due to deregulated expression of the HAS2 gene and/or alterations of HAS2 activity. The activity of HAS2 is regulated by post-translational modifications, including ubiquitination. In order to identify deubiquitinating enzymes (DUBs) that are involved in de-ubiquitination of HAS2, a complementary (cDNA) library of 69 Flag-HA-tagged human DUBs cloned into retroviral vectors was screened in human embryonic kidney (HEK) 293T cells for their ability to de-ubiquitinate myc-tagged HAS2. Several DUBs were found to decrease the ubiquitination of 6myc-HAS2, among which, the most effective were USP17 and USP4. USP17 efficiently removed polyubiquitination, whereas USP4 preferentially removed monoubiquitination of 6myc-HAS2. Co-immunoprecipitation studies revealed interactions between HAS2 and USP17, as well as between HAS2 and USP4, in membrane preparations of HEK293T cells. USP17 significantly stabilized 6myc-HAS2 protein levels, whereas USP4 did not. The silencing of USP17 led to decreased hyaluronan production, whereas the suppression of USP4 increased hyaluronan synthesis. Importantly, high levels of USP17 and HAS2 were detected in a panel of cancer cell lines compared to normal cells, and immunohistochemical stainings revealed higher expression of USP17 and HAS2 in tissues of lung cancer patients compared to normal tissue. In conclusion, USP17 and USP4 differently affect HAS2 ubiquitination, and the stability and function of HAS2.

Place, publisher, year, edition, pages
2017. Vol. 6, article id e348
Keywords [en]
cell cycle, ubiquitination, DUBs, growth, hyaluronan, cancer
National Category
Cell and Molecular Biology
Research subject
Biology with specialization in Molecular Biology; Biochemistry
Identifiers
URN: urn:nbn:se:uu:diva-312483DOI: 10.1038/oncsis.2017.45ISI: 000406047300003PubMedID: 28604766OAI: oai:DiVA.org:uu-312483DiVA, id: diva2:1064653
Funder
Swedish Cancer SocietyAvailable from: 2017-01-12 Created: 2017-01-10 Last updated: 2018-01-13Bibliographically approved
In thesis
1. Regulation of Hyaluronan Synthesis and Signaling via CD44 in Cancer
Open this publication in new window or tab >>Regulation of Hyaluronan Synthesis and Signaling via CD44 in Cancer
2017 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Hyaluronan is a ubiquitous glycosaminoglycan which is an important constituent of the extracellular matrix (ECM). In addition to organizing the extracellular matrix and regulating tissue homeostasis, hyaluronan, by binding to its main cell surface receptor CD44, is involved in intracellular signaling pathways regulating major cellular processes during development, wound healing, inflammation and cancer. Accumulation of hyaluronan in cancer promotes progression of the disease and correlates with poor prognosis. This thesis focuses on the regulation of hyaluronan synthesis and its signaling in normal and cancer cells.

Cancer cells in solid tumors are surrounded by stroma, which has an essential role in the growth and metastasis of tumors. Prominent members of the tumor stroma are fibroblasts, which synthesize ECM components, such as hyaluronan, and secrete growth factors, and activate intracellular signaling pathways. We demonstrate a cross-talk between the receptors for platelet-derived growth factor BB (PDGF-BB), transforming growth factor β (TGFβ) and CD44 in dermal fibroblasts. We found that PDGF-BB can activate the Smad signaling pathway downstream of the TGFβ receptor I (TβRI), and that PDGF-BB-induced migration depends on TβRI. CD44 forms a ternary complex with the receptors for PDGF-BB and TGFβ, and negatively regulates their signaling. Furthermore, we demonstrate that TGFβ stimulation of mammary epithelial cells transcriptionally upregulates hyaluronan synthase 2 (HAS2), which is essential for TGFβ-induced epithelial-mesenchymal transition (EMT); in this process, polarized epithelial cells adapt a mesenchymal phenotype which facilitates migration and invasion.

HAS2 protein activity and stability is regulated by posttranslational modifications, including ubiquitination. We investigated the ubiquitination of HAS2 in aggressive breast cancer cells, whose metastasizing capability depends on HAS2-synthesized hyaluronan. We identified two deubiquitinating enzymes, USP4 and USP17, which target HAS2 and affect its activity and stability.

In summary, these studies increase the knowledge about the regulation of hyaluronan production and its role in cancer progression.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2017. p. 54
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1291
Keywords
Hyaluronan, CD44, TGFβ, PDGF-BB, cancer, signaling, hyaluronan synthase, epithelial-mesenchymal transition
National Category
Cell and Molecular Biology
Research subject
Biology with specialization in Molecular Biology; Medical Science
Identifiers
urn:nbn:se:uu:diva-312485 (URN)978-91-554-9797-2 (ISBN)
Public defence
2017-03-03, B/B42, Biomedical Centre (BMC), Husargatan 3, Uppsala, 13:15 (English)
Opponent
Supervisors
Available from: 2017-02-10 Created: 2017-01-10 Last updated: 2018-01-13

Open Access in DiVA

fulltext(3959 kB)29 downloads
File information
File name FULLTEXT01.pdfFile size 3959 kBChecksum SHA-512
f87d0f70312ed0a2e747dfe72dba39b383d4bd72c0abb1588c22a1c8dee856c27d92e1ca2619d5bb1108f422365def94dc31ae06928a28bc33e961811e7b2c8b
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMed

Search in DiVA

By author/editor
Mehić, MerimaHebestreit, SandraHeldin, Carl-HenrikHeldin, Paraskevi
By organisation
Ludwig Institute for Cancer ResearchScience for Life Laboratory, SciLifeLabDepartment of Medical Biochemistry and Microbiology
In the same journal
Oncogenesis
Cell and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 29 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 526 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf