A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation

M.L. van de Weijer; M.C. Bassik; R.D. Luteijn; C.M. Voorburg; M.A. Lohuis; E. Kremmer; R.C. Hoeben; E.M. Leproust; S. Chen; H.M. Hoelen; M.E. Ressing; W. Patena; J.S. Weissman; M.T. McManus; E.J.H.J. Wiertz; R.J. Lebbink

doi:10.1038/ncomms4832

A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation

M.L. van de Weijer, M.C. Bassik, R.D. Luteijn, C.M. Voorburg, M.A. Lohuis, E. Kremmer, R.C. Hoeben, E.M. Leproust, S. Chen, H.M. Hoelen, M.E. Ressing, W. Patena, J.S. Weissman, M.T. McManus, E.J.H.J. Wiertz, R.J. Lebbink

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

Abstract

Misfolded ER proteins are retrotranslocated into the cytosol for degradation via the ubiquitin-proteasome system. The human cytomegalovirus protein US11 exploits this ER-associated protein degradation (ERAD) pathway to downregulate HLA class I molecules in virus-infected cells, thereby evading elimination by cytotoxic T-lymphocytes. US11-mediated degradation of HLA class I has been instrumental in the identification of key components of mammalian ERAD, including Derlin-1, p97, VIMP and SEL1L. Despite this, the process governing retrotranslocation of the substrate is still poorly understood. Here using a high-coverage genome-wide shRNA library, we identify the uncharacterized protein TMEM129 and the ubiquitin-conjugating E2 enzyme UBE2J2 to be essential for US11-mediated HLA class I downregulation. TMEM129 is an unconventional C4C4-type RING finger E3 ubiquitin ligase that resides within a complex containing various other ERAD components, including Derlin-1, Derlin-2, VIMP and p97, indicating that TMEM129 is an integral part of the ER-resident dislocation complex mediating US11-induced HLA class I degradation.

Original language	English
Pages (from-to)	3832
Number of pages	1
Journal	Nature Communications [E]
Volume	5
DOIs	https://doi.org/10.1038/ncomms4832
Publication status	Published - 2014

Keywords

Adenosine Triphosphatases
Cell Line, Tumor
Clustered Regularly Interspaced Short Palindromic Repeats
Cytomegalovirus
Cytomegalovirus Infections
Down-Regulation
Endoplasmic Reticulum
Endoplasmic Reticulum-Associated Degradation
HEK293 Cells
Histocompatibility Antigens Class I
Humans
Membrane Proteins
Nuclear Proteins
Protein Folding
Proteins
RNA Interference
RNA, Small Interfering
RNA-Binding Proteins
Selenoproteins
U937 Cells
Ubiquitin-Conjugating Enzymes
Ubiquitin-Protein Ligases
Viral Proteins
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

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10.1038/ncomms4832

http://www.ncbi.nlm.nih.gov/pubmed/24807418

Cite this

van de Weijer, M. L., Bassik, M. C., Luteijn, R. D., Voorburg, C. M., Lohuis, M. A., Kremmer, E., Hoeben, R. C., Leproust, E. M., Chen, S., Hoelen, H. M., Ressing, M. E., Patena, W., Weissman, J. S., McManus, M. T., Wiertz, E. J. H. J., & Lebbink, R. J. (2014). A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation. Nature Communications [E], 5, 3832. https://doi.org/10.1038/ncomms4832

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title = "A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation",

abstract = "Misfolded ER proteins are retrotranslocated into the cytosol for degradation via the ubiquitin-proteasome system. The human cytomegalovirus protein US11 exploits this ER-associated protein degradation (ERAD) pathway to downregulate HLA class I molecules in virus-infected cells, thereby evading elimination by cytotoxic T-lymphocytes. US11-mediated degradation of HLA class I has been instrumental in the identification of key components of mammalian ERAD, including Derlin-1, p97, VIMP and SEL1L. Despite this, the process governing retrotranslocation of the substrate is still poorly understood. Here using a high-coverage genome-wide shRNA library, we identify the uncharacterized protein TMEM129 and the ubiquitin-conjugating E2 enzyme UBE2J2 to be essential for US11-mediated HLA class I downregulation. TMEM129 is an unconventional C4C4-type RING finger E3 ubiquitin ligase that resides within a complex containing various other ERAD components, including Derlin-1, Derlin-2, VIMP and p97, indicating that TMEM129 is an integral part of the ER-resident dislocation complex mediating US11-induced HLA class I degradation.",

keywords = "Adenosine Triphosphatases, Cell Line, Tumor, Clustered Regularly Interspaced Short Palindromic Repeats, Cytomegalovirus, Cytomegalovirus Infections, Down-Regulation, Endoplasmic Reticulum, Endoplasmic Reticulum-Associated Degradation, HEK293 Cells, Histocompatibility Antigens Class I, Humans, Membrane Proteins, Nuclear Proteins, Protein Folding, Proteins, RNA Interference, RNA, Small Interfering, RNA-Binding Proteins, Selenoproteins, U937 Cells, Ubiquitin-Conjugating Enzymes, Ubiquitin-Protein Ligases, Viral Proteins, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't",

author = "{van de Weijer}, M.L. and M.C. Bassik and R.D. Luteijn and C.M. Voorburg and M.A. Lohuis and E. Kremmer and R.C. Hoeben and E.M. Leproust and S. Chen and H.M. Hoelen and M.E. Ressing and W. Patena and J.S. Weissman and M.T. McManus and E.J.H.J. Wiertz and R.J. Lebbink",

year = "2014",

doi = "10.1038/ncomms4832",

language = "English",

volume = "5",

pages = "3832",

journal = "Nature Communications [E]",

issn = "2041-1723",

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van de Weijer, ML, Bassik, MC, Luteijn, RD, Voorburg, CM, Lohuis, MA, Kremmer, E, Hoeben, RC, Leproust, EM, Chen, S, Hoelen, HM, Ressing, ME, Patena, W, Weissman, JS, McManus, MT, Wiertz, EJHJ & Lebbink, RJ 2014, 'A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation', Nature Communications [E], vol. 5, pp. 3832. https://doi.org/10.1038/ncomms4832

TY - JOUR

T1 - A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation

AU - van de Weijer, M.L.

AU - Bassik, M.C.

AU - Luteijn, R.D.

AU - Voorburg, C.M.

AU - Lohuis, M.A.

AU - Kremmer, E.

AU - Hoeben, R.C.

AU - Leproust, E.M.

AU - Chen, S.

AU - Hoelen, H.M.

AU - Ressing, M.E.

AU - Patena, W.

AU - Weissman, J.S.

AU - McManus, M.T.

AU - Wiertz, E.J.H.J.

AU - Lebbink, R.J.

PY - 2014

Y1 - 2014

N2 - Misfolded ER proteins are retrotranslocated into the cytosol for degradation via the ubiquitin-proteasome system. The human cytomegalovirus protein US11 exploits this ER-associated protein degradation (ERAD) pathway to downregulate HLA class I molecules in virus-infected cells, thereby evading elimination by cytotoxic T-lymphocytes. US11-mediated degradation of HLA class I has been instrumental in the identification of key components of mammalian ERAD, including Derlin-1, p97, VIMP and SEL1L. Despite this, the process governing retrotranslocation of the substrate is still poorly understood. Here using a high-coverage genome-wide shRNA library, we identify the uncharacterized protein TMEM129 and the ubiquitin-conjugating E2 enzyme UBE2J2 to be essential for US11-mediated HLA class I downregulation. TMEM129 is an unconventional C4C4-type RING finger E3 ubiquitin ligase that resides within a complex containing various other ERAD components, including Derlin-1, Derlin-2, VIMP and p97, indicating that TMEM129 is an integral part of the ER-resident dislocation complex mediating US11-induced HLA class I degradation.

AB - Misfolded ER proteins are retrotranslocated into the cytosol for degradation via the ubiquitin-proteasome system. The human cytomegalovirus protein US11 exploits this ER-associated protein degradation (ERAD) pathway to downregulate HLA class I molecules in virus-infected cells, thereby evading elimination by cytotoxic T-lymphocytes. US11-mediated degradation of HLA class I has been instrumental in the identification of key components of mammalian ERAD, including Derlin-1, p97, VIMP and SEL1L. Despite this, the process governing retrotranslocation of the substrate is still poorly understood. Here using a high-coverage genome-wide shRNA library, we identify the uncharacterized protein TMEM129 and the ubiquitin-conjugating E2 enzyme UBE2J2 to be essential for US11-mediated HLA class I downregulation. TMEM129 is an unconventional C4C4-type RING finger E3 ubiquitin ligase that resides within a complex containing various other ERAD components, including Derlin-1, Derlin-2, VIMP and p97, indicating that TMEM129 is an integral part of the ER-resident dislocation complex mediating US11-induced HLA class I degradation.

KW - Adenosine Triphosphatases

KW - Cell Line, Tumor

KW - Clustered Regularly Interspaced Short Palindromic Repeats

KW - Cytomegalovirus

KW - Cytomegalovirus Infections

KW - Down-Regulation

KW - Endoplasmic Reticulum

KW - Endoplasmic Reticulum-Associated Degradation

KW - HEK293 Cells

KW - Histocompatibility Antigens Class I

KW - Humans

KW - Membrane Proteins

KW - Nuclear Proteins

KW - Protein Folding

KW - Proteins

KW - RNA Interference

KW - RNA, Small Interfering

KW - RNA-Binding Proteins

KW - Selenoproteins

KW - U937 Cells

KW - Ubiquitin-Conjugating Enzymes

KW - Ubiquitin-Protein Ligases

KW - Viral Proteins

KW - Journal Article

KW - Research Support, N.I.H., Extramural

KW - Research Support, Non-U.S. Gov't

U2 - 10.1038/ncomms4832

DO - 10.1038/ncomms4832

M3 - Article

C2 - 24807418

SN - 2041-1723

VL - 5

SP - 3832

JO - Nature Communications [E]

JF - Nature Communications [E]

ER -

A high-coverage shRNA screen identifies TMEM129 as an E3 ligase involved in ER-associated protein degradation

Abstract

Keywords

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