Phosphorylation of the E3 ubiquitin protein ligase ITCH diminishes binding to its cognate E2 ubiquitin ligase

Heightened and extended inflammation underlies the pathogenesis of many disorders, including inflammatory bowel disease, sepsis, and inflammatory arthritis. Ubiquitin networks help dictate the strength and duration of inflammatory signaling. In innate immunity, the itchy E3 ubiquitin protein ligase...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:The Journal of biological chemistry 2018-01, Vol.293 (3), p.1100-1105
Hauptverfasser: Perez, Jessica M., Chen, Yinghua, Xiao, Tsan S., Abbott, Derek W.
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 1105
container_issue 3
container_start_page 1100
container_title The Journal of biological chemistry
container_volume 293
creator Perez, Jessica M.
Chen, Yinghua
Xiao, Tsan S.
Abbott, Derek W.
description Heightened and extended inflammation underlies the pathogenesis of many disorders, including inflammatory bowel disease, sepsis, and inflammatory arthritis. Ubiquitin networks help dictate the strength and duration of inflammatory signaling. In innate immunity, the itchy E3 ubiquitin protein ligase (ITCH)-A20 ubiquitin–editing complex inhibits receptor-interacting Ser/Thr kinase (RIPK) activation by removing Lys-63–linked polyubiquitinated chains from key proteins in the nuclear factor kappa B (NF-κB) signaling pathway. The complex then attaches polyubiquitinated chains to these proteins to target them for lysosomal or proteasomal destruction. ITCH is phosphorylated and thereby inhibited by inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) to fine-tune the inflammatory response to the strength of the offending signal. However, the biochemical mechanism by which E3 ubiquitination is impaired by IKK-driven phosphorylation remains unclear. Here, we report that this phosphorylation impedes ITCH binding to its cognate E2 ubiquitin ligase, UbcH7. Using CRISPR-Cas9 genetic knockout to mimic the ITCH-UbcH7–inhibited state, we further show that genetic UbcH7 deficiency phenocopies ITCH phosphorylation in regulating RIPK2 ubiquitination. We conclude that phosphorylation can disrupt the binding of an E3 ubiquitin ligase to an E2-conjugating enzyme, leading to prolonged inflammatory signaling. To our knowledge, this is the first report of E3 ubiquitin ligase phosphorylation inhibiting E3 ligase activity by impairing E2–E3 complex formation.
doi_str_mv 10.1074/jbc.RA117.000408
format Article
fullrecord <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5777250</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0021925820396794</els_id><sourcerecordid>1974007701</sourcerecordid><originalsourceid>FETCH-LOGICAL-c447t-8f6d21c451b830e2f3ccdd335c83acdf8843e2c823868518b063a585682d00a63</originalsourceid><addsrcrecordid>eNp1kUFv1DAQRi0Eokvhzgn5yCXL2E5iLwekqmqhUiUQAomb5djOZqrE3tpOpf77pqStyoHTHPx9b0Z-hLxnsGUg609Xnd3-PGFMbgGgBvWCbBgoUYmG_XlJNgCcVTveqCPyJucruA_t2GtyxHeccQnthkw_hpgPQ0y3oykYA409LYOnZ4LOHV7PWDDQQ4rFL3PEvcmeXvw6_UYdThgwDz7TDoPDsKclUiyZ2rgPpiwI_gyxVt-SV70Zs3_3MI_J7_OzhVZdfv96cXpyWdm6lqVSfes4s3XDOiXA815Y65wQjVXCWNcrVQvPreJCtaphqoNWmEY1reIOwLTimHxZuYe5m7yzPpRkRn1IOJl0q6NB_e9LwEHv441upJS8gQXw8QGQ4vXsc9ETZuvH0QQf56zZTtYAUgJborBGbYo5J98_rWGg7y3pxZL-a0mvlpbKh-fnPRUetSyBz2vAL590gz7pbNEH6x0mb4t2Ef9PvwPXxaLx</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1974007701</pqid></control><display><type>article</type><title>Phosphorylation of the E3 ubiquitin protein ligase ITCH diminishes binding to its cognate E2 ubiquitin ligase</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><creator>Perez, Jessica M. ; Chen, Yinghua ; Xiao, Tsan S. ; Abbott, Derek W.</creator><creatorcontrib>Perez, Jessica M. ; Chen, Yinghua ; Xiao, Tsan S. ; Abbott, Derek W.</creatorcontrib><description>Heightened and extended inflammation underlies the pathogenesis of many disorders, including inflammatory bowel disease, sepsis, and inflammatory arthritis. Ubiquitin networks help dictate the strength and duration of inflammatory signaling. In innate immunity, the itchy E3 ubiquitin protein ligase (ITCH)-A20 ubiquitin–editing complex inhibits receptor-interacting Ser/Thr kinase (RIPK) activation by removing Lys-63–linked polyubiquitinated chains from key proteins in the nuclear factor kappa B (NF-κB) signaling pathway. The complex then attaches polyubiquitinated chains to these proteins to target them for lysosomal or proteasomal destruction. ITCH is phosphorylated and thereby inhibited by inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) to fine-tune the inflammatory response to the strength of the offending signal. However, the biochemical mechanism by which E3 ubiquitination is impaired by IKK-driven phosphorylation remains unclear. Here, we report that this phosphorylation impedes ITCH binding to its cognate E2 ubiquitin ligase, UbcH7. Using CRISPR-Cas9 genetic knockout to mimic the ITCH-UbcH7–inhibited state, we further show that genetic UbcH7 deficiency phenocopies ITCH phosphorylation in regulating RIPK2 ubiquitination. We conclude that phosphorylation can disrupt the binding of an E3 ubiquitin ligase to an E2-conjugating enzyme, leading to prolonged inflammatory signaling. To our knowledge, this is the first report of E3 ubiquitin ligase phosphorylation inhibiting E3 ligase activity by impairing E2–E3 complex formation.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.RA117.000408</identifier><identifier>PMID: 29212706</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>E3 ubiquitin ligase ; HEK293 Cells ; Humans ; IKKβ ; Immunoprecipitation ; inflammation ; Inflammation - metabolism ; ITCH ; NF-kappa B ; NF-kappa B - metabolism ; NF-κB ; Phosphorylation ; Protein Binding ; Repressor Proteins - metabolism ; Signal Transduction ; UbcH7 ; ubiquitin ligase ; Ubiquitin-Protein Ligases - metabolism ; ubiquitination ; Ubiquitination - physiology</subject><ispartof>The Journal of biological chemistry, 2018-01, Vol.293 (3), p.1100-1105</ispartof><rights>2018 © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.</rights><rights>2018 by The American Society for Biochemistry and Molecular Biology, Inc.</rights><rights>2018 by The American Society for Biochemistry and Molecular Biology, Inc. 2018 The American Society for Biochemistry and Molecular Biology, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c447t-8f6d21c451b830e2f3ccdd335c83acdf8843e2c823868518b063a585682d00a63</citedby><cites>FETCH-LOGICAL-c447t-8f6d21c451b830e2f3ccdd335c83acdf8843e2c823868518b063a585682d00a63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5777250/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5777250/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29212706$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Perez, Jessica M.</creatorcontrib><creatorcontrib>Chen, Yinghua</creatorcontrib><creatorcontrib>Xiao, Tsan S.</creatorcontrib><creatorcontrib>Abbott, Derek W.</creatorcontrib><title>Phosphorylation of the E3 ubiquitin protein ligase ITCH diminishes binding to its cognate E2 ubiquitin ligase</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>Heightened and extended inflammation underlies the pathogenesis of many disorders, including inflammatory bowel disease, sepsis, and inflammatory arthritis. Ubiquitin networks help dictate the strength and duration of inflammatory signaling. In innate immunity, the itchy E3 ubiquitin protein ligase (ITCH)-A20 ubiquitin–editing complex inhibits receptor-interacting Ser/Thr kinase (RIPK) activation by removing Lys-63–linked polyubiquitinated chains from key proteins in the nuclear factor kappa B (NF-κB) signaling pathway. The complex then attaches polyubiquitinated chains to these proteins to target them for lysosomal or proteasomal destruction. ITCH is phosphorylated and thereby inhibited by inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) to fine-tune the inflammatory response to the strength of the offending signal. However, the biochemical mechanism by which E3 ubiquitination is impaired by IKK-driven phosphorylation remains unclear. Here, we report that this phosphorylation impedes ITCH binding to its cognate E2 ubiquitin ligase, UbcH7. Using CRISPR-Cas9 genetic knockout to mimic the ITCH-UbcH7–inhibited state, we further show that genetic UbcH7 deficiency phenocopies ITCH phosphorylation in regulating RIPK2 ubiquitination. We conclude that phosphorylation can disrupt the binding of an E3 ubiquitin ligase to an E2-conjugating enzyme, leading to prolonged inflammatory signaling. To our knowledge, this is the first report of E3 ubiquitin ligase phosphorylation inhibiting E3 ligase activity by impairing E2–E3 complex formation.</description><subject>E3 ubiquitin ligase</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>IKKβ</subject><subject>Immunoprecipitation</subject><subject>inflammation</subject><subject>Inflammation - metabolism</subject><subject>ITCH</subject><subject>NF-kappa B</subject><subject>NF-kappa B - metabolism</subject><subject>NF-κB</subject><subject>Phosphorylation</subject><subject>Protein Binding</subject><subject>Repressor Proteins - metabolism</subject><subject>Signal Transduction</subject><subject>UbcH7</subject><subject>ubiquitin ligase</subject><subject>Ubiquitin-Protein Ligases - metabolism</subject><subject>ubiquitination</subject><subject>Ubiquitination - physiology</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kUFv1DAQRi0Eokvhzgn5yCXL2E5iLwekqmqhUiUQAomb5djOZqrE3tpOpf77pqStyoHTHPx9b0Z-hLxnsGUg609Xnd3-PGFMbgGgBvWCbBgoUYmG_XlJNgCcVTveqCPyJucruA_t2GtyxHeccQnthkw_hpgPQ0y3oykYA409LYOnZ4LOHV7PWDDQQ4rFL3PEvcmeXvw6_UYdThgwDz7TDoPDsKclUiyZ2rgPpiwI_gyxVt-SV70Zs3_3MI_J7_OzhVZdfv96cXpyWdm6lqVSfes4s3XDOiXA815Y65wQjVXCWNcrVQvPreJCtaphqoNWmEY1reIOwLTimHxZuYe5m7yzPpRkRn1IOJl0q6NB_e9LwEHv441upJS8gQXw8QGQ4vXsc9ETZuvH0QQf56zZTtYAUgJborBGbYo5J98_rWGg7y3pxZL-a0mvlpbKh-fnPRUetSyBz2vAL590gz7pbNEH6x0mb4t2Ef9PvwPXxaLx</recordid><startdate>20180119</startdate><enddate>20180119</enddate><creator>Perez, Jessica M.</creator><creator>Chen, Yinghua</creator><creator>Xiao, Tsan S.</creator><creator>Abbott, Derek W.</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20180119</creationdate><title>Phosphorylation of the E3 ubiquitin protein ligase ITCH diminishes binding to its cognate E2 ubiquitin ligase</title><author>Perez, Jessica M. ; Chen, Yinghua ; Xiao, Tsan S. ; Abbott, Derek W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c447t-8f6d21c451b830e2f3ccdd335c83acdf8843e2c823868518b063a585682d00a63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>E3 ubiquitin ligase</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>IKKβ</topic><topic>Immunoprecipitation</topic><topic>inflammation</topic><topic>Inflammation - metabolism</topic><topic>ITCH</topic><topic>NF-kappa B</topic><topic>NF-kappa B - metabolism</topic><topic>NF-κB</topic><topic>Phosphorylation</topic><topic>Protein Binding</topic><topic>Repressor Proteins - metabolism</topic><topic>Signal Transduction</topic><topic>UbcH7</topic><topic>ubiquitin ligase</topic><topic>Ubiquitin-Protein Ligases - metabolism</topic><topic>ubiquitination</topic><topic>Ubiquitination - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Perez, Jessica M.</creatorcontrib><creatorcontrib>Chen, Yinghua</creatorcontrib><creatorcontrib>Xiao, Tsan S.</creatorcontrib><creatorcontrib>Abbott, Derek W.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Perez, Jessica M.</au><au>Chen, Yinghua</au><au>Xiao, Tsan S.</au><au>Abbott, Derek W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Phosphorylation of the E3 ubiquitin protein ligase ITCH diminishes binding to its cognate E2 ubiquitin ligase</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2018-01-19</date><risdate>2018</risdate><volume>293</volume><issue>3</issue><spage>1100</spage><epage>1105</epage><pages>1100-1105</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Heightened and extended inflammation underlies the pathogenesis of many disorders, including inflammatory bowel disease, sepsis, and inflammatory arthritis. Ubiquitin networks help dictate the strength and duration of inflammatory signaling. In innate immunity, the itchy E3 ubiquitin protein ligase (ITCH)-A20 ubiquitin–editing complex inhibits receptor-interacting Ser/Thr kinase (RIPK) activation by removing Lys-63–linked polyubiquitinated chains from key proteins in the nuclear factor kappa B (NF-κB) signaling pathway. The complex then attaches polyubiquitinated chains to these proteins to target them for lysosomal or proteasomal destruction. ITCH is phosphorylated and thereby inhibited by inhibitor of nuclear factor kappa B kinase subunit beta (IKKβ) to fine-tune the inflammatory response to the strength of the offending signal. However, the biochemical mechanism by which E3 ubiquitination is impaired by IKK-driven phosphorylation remains unclear. Here, we report that this phosphorylation impedes ITCH binding to its cognate E2 ubiquitin ligase, UbcH7. Using CRISPR-Cas9 genetic knockout to mimic the ITCH-UbcH7–inhibited state, we further show that genetic UbcH7 deficiency phenocopies ITCH phosphorylation in regulating RIPK2 ubiquitination. We conclude that phosphorylation can disrupt the binding of an E3 ubiquitin ligase to an E2-conjugating enzyme, leading to prolonged inflammatory signaling. To our knowledge, this is the first report of E3 ubiquitin ligase phosphorylation inhibiting E3 ligase activity by impairing E2–E3 complex formation.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>29212706</pmid><doi>10.1074/jbc.RA117.000408</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record>
fulltext fulltext
identifier ISSN: 0021-9258
ispartof The Journal of biological chemistry, 2018-01, Vol.293 (3), p.1100-1105
issn 0021-9258
1083-351X
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5777250
source MEDLINE; EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection
subjects E3 ubiquitin ligase
HEK293 Cells
Humans
IKKβ
Immunoprecipitation
inflammation
Inflammation - metabolism
ITCH
NF-kappa B
NF-kappa B - metabolism
NF-κB
Phosphorylation
Protein Binding
Repressor Proteins - metabolism
Signal Transduction
UbcH7
ubiquitin ligase
Ubiquitin-Protein Ligases - metabolism
ubiquitination
Ubiquitination - physiology
title Phosphorylation of the E3 ubiquitin protein ligase ITCH diminishes binding to its cognate E2 ubiquitin ligase
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T14%3A58%3A27IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Phosphorylation%20of%20the%20E3%20ubiquitin%20protein%20ligase%20ITCH%20diminishes%20binding%20to%20its%20cognate%20E2%20ubiquitin%20ligase&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Perez,%20Jessica%20M.&rft.date=2018-01-19&rft.volume=293&rft.issue=3&rft.spage=1100&rft.epage=1105&rft.pages=1100-1105&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.RA117.000408&rft_dat=%3Cproquest_pubme%3E1974007701%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1974007701&rft_id=info:pmid/29212706&rft_els_id=S0021925820396794&rfr_iscdi=true