Rhomboid protease RHBDL4 promotes retrotranslocation of aggregation-prone proteins for degradation

Bock, Josephine; Kühnle, Nathalie; Knopf, Julia D.; Landscheidt, Nina; Lee, Jingu; Ye, Yihong; Lemberg, Marius K.

doi:10.1016/j.celrep.2022.111175

Cited by 16 publications

(13 citation statements)

References 61 publications

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“…Although they were initially discovered as intramembrane proteases, rhomboid proteases are able to cleave substrates both in the plane of the membrane as well as in their ectodomains (Avci & Lemberg, 2015; Bock et al , 2022; Paschkowsky et al ., 2016; Recinto et al ., 2018; Strisovsky et al ., 2009). Here, we demonstrate that RHBDL4-mediated cleavages of APP in the ER generate Aη-like peptides.…”

Section: Discussionmentioning

confidence: 99%

Eta-secretase-like processing of the amyloid precursor protein (APP) by RHBDL4

Ylauna Christine Megane,

Paschkowsky,

Recinto

et al. 2023

Preprint

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The amyloid precursor protein (APP) has been extensively studied with regards to its contribution to the pathology of Alzheimer's disease. APP is an ubiquitously expressed type I transmembrane protein synthesized in the endoplasmic reticulum (ER) and translocated to the plasma membrane where it undergoes proteolytic cleavages by several identified proteases. Conversely to other known proteases, we previously elucidated human rhomboid protease RHBDL4 as a novel APP processing enzyme where several cleavages likely occur already in the ER. Interestingly, the pattern of RHBDL4-derived large APP C-terminal fragments resemble those generated by the η-secretase or MT5-MMP, which was described to generate so called Aη fragments. The similarity in large APP C-terminal fragments between both proteases raised the question whether RHBDL4 may contribute to η-secretase activity and Aη-like fragments. Here, we identified two cleavage sites of RHBDL4 in APP by mass spectrometry, which, intriguingly, lie in close proximity to the cleavage sites of MT5-MMP. Indeed, we observed that RHBDL4 generates Aη-like fragments in vitro without contributions of α-, β-, or γ-secretases. Such Aη-like fragments are likely generated in the ER since RHBDL4-derived APP-C-terminal fragments do not reach the cell surface. Inherited, familial APP mutations appear to not affect this processing pathway. In RHBDL4 knockout mice, we observed increased cerebral full length APP levels in comparison to WT brains in support of RHBDL4 being a physiologically relevant protease for APP. Furthermore, we found secreted Aη fragments in dissociated mixed cortical cultures from wild type mice, however significantly less Aη fragments in cultures from RHBDL4 knockout mice. Our data underscores that RHBDL4 contributes to η-secretease-like processing of APP and that RHBDL4 is a physiologically relevant protease for APP.

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Section: Discussionmentioning

confidence: 99%

Eta-secretase-like processing of the amyloid precursor protein (APP) by RHBDL4

Ylauna Christine Megane,

Paschkowsky,

Recinto

et al. 2023

Preprint

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“…Further overlap between our data sets exists for VCP/p97 (TERA), VRK2, MAN1, and COPG2 in relation to our stringent analysis. In addition, protein interactors were investigated with pull-down assays by the Lemberg lab, who discovered, for example, protein disulfide isomerases (PDIs), 9 which we could confirm in our BioID assay as well (PDIA6, see the Supporting Information). Conversely, there are also remarkable differences between our BioID analyses and those of Ikeda and Freeman.…”

Section: ■ Discussionmentioning

confidence: 87%

“…In addition, cancer-relevant signaling pathways including AKT and β-catenin were found to be altered when RHBDL4 expression was increased. ,, RHBDL4 has been shown to play a role in the endoplasmic reticulum (ER)-associated degradation (ERAD), where it facilitates the retro-translocation process of misfolded proteins . The globular C-terminus of RHBDL4 contains a ubiquitin-interacting motif and an interaction site for the transitional ER ATPase, VCP/p97, which is known to pull misfolded proteins out of the ER to subsequently direct them to the proteasome. , Thus, it was proposed that RHBDL4 may cleave misfolded or aggregation-prone proteins that are stalled in the retro-translocation process . A number of substrates have been identified for RHBDL4, including Bcl-2-interacting killer (BIK), oligosaccharyl transferase (OST), pre T-cell receptor substrate alpha (pTα), sterol regulatory element-binding protein 1 (SREBP1), and tumor suppressor-activated pathway-6 (TSAP6). ,,− A recent paper identified dozens more RHBDL4 substrates, and among many of them, RHBDL4 appears to control the removal of C-terminal ER-retention motifs of ER chaperones .…”

Section: Introductionmentioning

confidence: 99%

“…7,8 Thus, it was proposed that RHBDL4 may cleave misfolded or aggregationprone proteins that are stalled in the retro-translocation process. 9 A number of substrates have been identified for RHBDL4, including Bcl-2-interacting killer (BIK), oligosaccharyl transferase (OST), pre T-cell receptor substrate alpha (pTα), sterol regulatory element-binding protein 1 (SREBP1), and tumor suppressor-activated pathway-6 (TSAP6). 6,7,10−12 A recent paper identified dozens more RHBDL4 substrates, and among many of them, RHBDL4 appears to control the removal of C-terminal ER-retention motifs of ER chaperones.…”

Section: ■ Introductionmentioning

confidence: 99%

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Putative Protein Interactome of the Rhomboid Protease RHBDL4

Hsiao

Penalva

et al. 2023

Biochemistry

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The physiological functions of the rhomboid-related protein 4 (RHBDL4) are emerging, but their molecular details remain unclear. Because increased expression of RHBDL4 has been clinically linked to poorer outcomes in cancer patients, this association urgently demands a better understanding of RHBDL4. To elucidate the molecular interactions and pathways that RHBDL4 may be involved in, we conducted proximity-dependent biotin identification (BioID) assays. Our analyses corroborated several of the expected protein interactors such as the transitional endoplasmic reticulum (ER) ATPase VCP/p97 (TERA), but they also described novel putative interactors including IRS4, PGAM5, and GORS2. Using proximity-ligation assays, we validated VCP/ p97, COPB, and VRK2 as proteins that are in proximity to RHBDL4. Overall, our results support the emerging functions of RHBDL4 in ER quality control and also point toward putative RHBDL4 functions in protein membrane insertion and membrane organization and trafficking.

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“…This study is the first to demonstrate chaperone-like activity for any rhomboid protein. Many rhomboid proteins use similar functions to Dfm1 to promote retrotranslocation, and the rhomboid protease RHBDL4 has recently been characterized as acting on aggregation-prone substrates [19,28,[35][36][37]. It will be an interesting and important further line of inquiry to determine if a chaperone-like activity is a common ability of rhomboid proteins, both for the pseudoproteases and proteases.…”

Section: Introductionmentioning

confidence: 99%

Yeast derlin Dfm1 employs a chaperone-like function to resolve misfolded membrane protein stress

et al. 2023

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Protein aggregates are a common feature of diseased and aged cells. Membrane proteins comprise a quarter of the proteome, and yet, it is not well understood how aggregation of membrane proteins is regulated and what effects these aggregates can have on cellular health. We have determined in yeast that the derlin Dfm1 has a chaperone-like activity that influences misfolded membrane protein aggregation. We establish that this function of Dfm1 does not require recruitment of the ATPase Cdc48 and it is distinct from Dfm1’s previously identified function in dislocating misfolded membrane proteins from the endoplasmic reticulum (ER) to the cytosol for degradation. Additionally, we assess the cellular impacts of misfolded membrane proteins in the absence of Dfm1 and determine that misfolded membrane proteins are toxic to cells in the absence of Dfm1 and cause disruptions to proteasomal and ubiquitin homeostasis.

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Rhomboid protease RHBDL4 promotes retrotranslocation of aggregation-prone proteins for degradation

Cited by 16 publications

References 61 publications

Eta-secretase-like processing of the amyloid precursor protein (APP) by RHBDL4

Eta-secretase-like processing of the amyloid precursor protein (APP) by RHBDL4

Putative Protein Interactome of the Rhomboid Protease RHBDL4

Yeast derlin Dfm1 employs a chaperone-like function to resolve misfolded membrane protein stress

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