The Evolutionary Conserved Transmembrane BAX Inhibitor Motif (TMBIM) Containing Protein Family is Essential for the Development and Survival of Drosophila Melanogaster

The Evolutionary Conserved Transmembrane BAX Inhibitor Motif (TMBIM) Containing Protein Family is Essential for the Development and Survival of Drosophila Melanogaster | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The Evolutionary Conserved Transmembrane BAX Inhibitor Motif (TMBIM) Containing Protein Family is Essential for the Development and Survival of Drosophila Melanogaster Li Zhang, Sebastian Buhr, Vibha Prasad, Aaron Voigt, Axel Methner This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-113374/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 02 Sep, 2021 Read the published version in Frontiers in Cell and Developmental Biology → Version 1 posted You are reading this latest preprint version Abstract The Transmembrane BAX Inhibitor Motif (TMBIM) protein family consists of six evolutionarily conserved hydrophobic proteins that affect programmed cell death and the regulation of intracellular calcium levels. The bacterial orthologue BsYetJ is a pH-dependent calcium channel. We here identified six TMBIM family members in Drosophila melanogaster and studied the effect of their RNAi-mediated knockdown using ubiquitous and tissue-specific drivers. Mammalian TMBIM6 and TMBIM5 have obvious orthologs while this is more dubious for the other family members. Ubiquitous knockdown of family members dmTMBIM1,4,5, and 6 caused failed eclosing and tissue-specific knockdown resulted in a dramatically decreased lifespan. On the contrary, knockdown of dmTMBIM3, surprisingly, extended lifespan. Only knockdown of dmTMBIM6 increased the ER calcium levels of Pdf neurons. Neural knockdown of dmTMBIM2,3, and 4 increased ER stress, as indicated by increased Xbp1 splicing. Interestingly, TMBIM1 and TMBIM6 have a very similar expression pattern and their knockdown phenocopied each other. Also, knockdown of TMBIM1 resulted in upregulation of TMBIM6 and vice versa further suggesting a genetic interaction between these two genes. Our data demonstrate that most TMBIM proteins are essential for fly development and survival but, despite their shared protein structure, affect cell survival through different mechanisms. Cell Survival and Cell Death General Cell Biology & Physiology RECS1 PP1201 LFG3 MST100 MSTP100 NMP35 LFG FAIM2 LFG2 NGP35 KIAA0950 LFG1 HNRGW NMDARA1 MGC99687 GRINA GBP GAAP S1R LFG4 ZPRO CGI-119 GHITM MICS1 DERP2 My021 PTD010 HSPC282 FLJ26584 DKFZp566C0746 Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Full Text Supplementary Files SupplementaryInformation.docx Supplemental Figure S1 Expression levels of CG9722 (dmTMBIM1) in the CG9722 RNAi#2 line and MiMIC line a) tub>>CG9722-RNAi#2 (BL6679) showed increased transcriptional levels of CG9722 assessed by quantitative qPCR in whole adults. b) CG9722-MiMIC line (BL44979), which is supposed to be a CG9722 knockout does not alter transcriptional levels of CG9722. c) Location of primers in respect to the CG9722 MiMIC insertion cassette. In homozygous CG9722 MiMIC flies, PCR using primer pair 1 amplifies a product of over 500 bp indicating no insertion of the MiMIC cassette. Primer pair 2 suggests the existence of the MiMIC cassette in the genome. Primer pairs 1for/2rev and 2for/1rev respectively 1for/2for and 1rev/2rev did not result in any PCR product in line with the lack of the MIMIC cassette within the CG9722 locus. Data are represented as mean ± SD. Unpaired t-test were performed using GraphPad Prism. *p<0.05. Supplemental Table S1 RNAi lines used in this study. Cite Share Download PDF Status: Published Journal Publication published 02 Sep, 2021 Read the published version in Frontiers in Cell and Developmental Biology → Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-113374","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":5419270,"identity":"f4f0df7d-11f3-41d2-8cab-9616be5866cb","order_by":0,"name":"Li Zhang","email":"","orcid":"","institution":"University Medical Center of the Johannes Gutenberg University Mainz","correspondingAuthor":false,"prefix":"","firstName":"Li","middleName":"","lastName":"Zhang","suffix":""},{"id":5419271,"identity":"af55dd82-5538-4acc-bfad-2f6b5f39a020","order_by":1,"name":"Sebastian Buhr","email":"","orcid":"","institution":"University Medical Center of the Johannes Gutenberg University Mainz","correspondingAuthor":false,"prefix":"","firstName":"Sebastian","middleName":"","lastName":"Buhr","suffix":""},{"id":5419272,"identity":"ada2399e-28c6-455b-9d56-f639490d69c9","order_by":2,"name":"Vibha Prasad","email":"","orcid":"","institution":"Universitätsklinikum Aachen","correspondingAuthor":false,"prefix":"","firstName":"Vibha","middleName":"","lastName":"Prasad","suffix":""},{"id":5419273,"identity":"a6d45895-6164-49fd-8283-b5a170445c96","order_by":3,"name":"Aaron Voigt","email":"","orcid":"","institution":"Universitätsklinikum Aachen","correspondingAuthor":false,"prefix":"","firstName":"Aaron","middleName":"","lastName":"Voigt","suffix":""},{"id":5419274,"identity":"73871272-92d1-4b27-b088-113827d85853","order_by":4,"name":"Axel Methner","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA80lEQVRIiWNgGAWjYHACxgMg0gBEfABiNnYi9IC0SIC0MM4AaWEmRQszD4hLSAv/jOQDBxhzbOrMpZuPfbb5tU2ej5mB8cPHHNxaJM4cSzjAuC1NwnLOseTZuX23DduYGZglZ27DY83xHgOglsMSBjdyjJlze24zArWwMfPi0SJ/mP8DUMt/oJb8z8yWPbftCWoxON7DANRyAGQLMzPDj9uJBLUYnjlmcCBxW7Lkhhtpxoy9DbeT25gZm/H6Re5G8sMHH7fZ8RvcSH7M8OPPbdv57c0HP3zE530QSIAxGNvAZAMB9SjgDymKR8EoGAWjYKQAAKZTUnJziOTDAAAAAElFTkSuQmCC","orcid":"","institution":"University Medical Center of the Johannes Gutenberg University Mainz","correspondingAuthor":true,"prefix":"","firstName":"Axel","middleName":"","lastName":"Methner","suffix":""}],"badges":[],"createdAt":"2020-11-21 16:59:01","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-113374/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-113374/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.3389/fcell.2021.666484","type":"published","date":"2021-09-03T00:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":3985219,"identity":"a03e9840-fdd8-46e2-8ecc-207a40990c27","added_by":"auto","created_at":"2020-12-03 15:10:11","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":38734,"visible":true,"origin":"","legend":"Phylogenetic relationship of fly and human TMBIM proteins\nPhylogenetic analysis of the sequence alignment of human and Drosophila TMBIM proteins identifies CG2076 and CG1287 as orthologues for TMBIM5 and CG7188 as TMBIM6, TMBIM4\nis most probably CG30370 while the relationship of TMBIM1-3 to their fly orthologues is less clear. The bacterial homologue BsYetJ is most closely related to TMBIM5 and TMBIM6. The\nbranch support values are indicated above the branches. The phylogenetic tree was created\nusing the Maximum Likelihood algorithm with the BLOSUM62 substitution matrix and 100\nbootstrap trials. The dashed lines indicate closest homologues according to BLASTP.","description":"","filename":"Fig1.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/d5c7191b2872e742ea1a3f7a.JPG"},{"id":3985221,"identity":"f991a48a-3cea-48fd-9d7a-c4888e968a01","added_by":"auto","created_at":"2020-12-03 15:10:12","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":155955,"visible":true,"origin":"","legend":"Expression levels and phenotype of CG7188 (dmTMBIM6) knockdown in flies\na) Temporal and tissue expression of CG7188/dmTMBIM5 from Flybase (http://flybase.org/).\nb) Viability of knockdown (KD) with different GAL4 drivers and the knockdown efficiency of two UAS-CG7188-RNAi lines (#1, #2) assessed by quantitative PCR in whole larvae of\nMef2\u003e\u003eCG7188-RNAi. c) Effects of neural KD by elav-GAL4 of CG7188 on eclosing, wing\nphenotype and life span. d) Crawling ability of larvae of muscular KD (Mef2\u003e\u003eCG7188-RNAi).\ne) Activity of Pdf\u003e\u003eCG7188-RNA under 12/12 hours light/dark cycle. Data are represented as\nmean ± SD. One-way ANOVA followed by Newman-Keul’s multiple comparison test for\nmultiple comparison and un-paired t-test for two groups comparison were performed using\nGraphPad Prism. *p\u003c0.05.","description":"","filename":"Fig2.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/e76602d92ba0f6957fe09b7d.JPG"},{"id":3985222,"identity":"23af553a-08ed-448c-acfa-503dab00ecc2","added_by":"auto","created_at":"2020-12-03 15:10:12","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":159838,"visible":true,"origin":"","legend":"Expression levels and phenotype of CG2076 (dmTMBIM5) knockdown in flies\na) Temporal and tissue expression of CG2076/dmTMBIM5 from Flybase (http://flybase.org/).\nb) Viability of CG2076 knockdown (KD) flies by different GAL4 drivers and the KD efficiency\nof UAS-CG2076-RNAi (#1, #2) assessed by quantitative PCR in whole larvae of\nMef2\u003e\u003eCG2076-RNAi. c) Eclosing rate of pupa and lifespan of elav\u003e\u003eCG2076-RNAi. d)\nCrawling ability of larvae, climbing ability and lifespan of adults of Mef2\u003e\u003eCG2076-RNAi. e)\nActivity of Pdf\u003e\u003eCG2076-RNAi flies under 12/12 hours light/dark cycle. Data are represented\nas mean ± SD. One-way ANOVA followed by Newman-Keul’s multiple comparison test for\nmultiple comparison and two-way ANOVA followed by Sidak’s multiple comparison test for multiple comparison under two independent conditions were performed using GraphPad\nPrism. *p\u003c0.05.","description":"","filename":"Fig3.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/44fcacc8909be77ab03fe4ae.JPG"},{"id":3985223,"identity":"a62a41f5-6d82-4b93-bce2-b041cba0d899","added_by":"auto","created_at":"2020-12-03 15:10:12","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":148177,"visible":true,"origin":"","legend":"Expression levels and phenotype of CG30379 (putative dmTMBIM4)\nknockdown in flies\na) Temporal and tissue expression of CG30379/dmTMBIM4 from Flybase (http://flybase.org/).\nb) Viability of CG30379 knockdown (KD) flies by different GAL4 drivers and the KD efficiency\nof UAS-CG30379-RNAi (#1, #2) assessed by quantitative PCR in fly heads of\n17 elav\u003e\u003eCG30379-RNAi (#1) and elav\u003e\u003eCG30379-RNAi (#2). c) Eclosing rate of pupa and lifespan of elav\u003e\u003eCG30379-RNAi. d) Crawling ability of larvae, climbing ability and lifespan of adults of Mef2\u003e\u003eCG30379-RNAi. e) Activity of Pdf\u003e\u003eCG30379-RNAi flies under 12/12 hours light/dark cycle. Data are represented as mean ± SD. One-way ANOVA followed by Newman-Keul’s multiple comparison test for multiple comparison and two-way ANOVA followed by\nSidak’s multiple comparison test for multiple comparison under two independent conditions\nwere performed using GraphPad Prism. *p\u003c0.05.","description":"","filename":"Fig4.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/b21c55b1e52da5495c7c7280.JPG"},{"id":3985224,"identity":"89767336-6f01-4f80-bf75-ff29d0c69b4f","added_by":"auto","created_at":"2020-12-03 15:10:12","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":155303,"visible":true,"origin":"","legend":"Expression levels and phenotype of CG3798 (putative dmTMBIM3) knockdown\nin flies a) Temporal and tissue expression of CG3798/dmTMBIM3 from Flybase (http://flybase.org/).\nb) Viability of CG3798 knockdown (KD) flies by different GAL4 drivers and the KD efficiency\nof UAS-CG3798-RNAi (#1, #2) assessed by quantitative PCR in whole adults of\ntub\u003e\u003eCG3798-RNAi. c) Eclosing rate of pupa and lifespan of elav\u003e\u003eCG3798-RNAi. d)\nCrawling ability of larvae, climbing ability and lifespan of adults of Mef2\u003e\u003eCG3798-RNAi. e)\nActivity of Pdf\u003e\u003eCG3798-RNAi flies under 12/12 hours light/dark cycle. Data are represented\nas mean ± SD. One-way ANOVA followed by Newman-Keul’s multiple comparison test for\nmultiple comparison and two-way ANOVA followed by Sidak’s multiple comparison test for multiple comparison under two independent conditions were performed using GraphPad\nPrism. *p\u003c0.05.","description":"","filename":"Fig5.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/d0fbed163a3c735f2e4a8fd6.JPG"},{"id":3985225,"identity":"80d55711-73e0-471a-8309-d20f11f42250","added_by":"auto","created_at":"2020-12-03 15:10:13","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":153275,"visible":true,"origin":"","legend":"Expression levels and phenotype of CG3814 (putative dmTMBIM2) knockdown\nin flies a) Temporal and tissue expression of CG3814/dmTMBIM2 from Flybase (http://flybase.org/).\nb) Viability of CG3814 knockdown (KD) flies by different GAL4 drivers and the knockdown\nefficiency of UAS- CG3814-RNAi (#1, #2) assessed by quantitative PCR in whole adults of\ntub\u003e\u003eCG3814-RNAi. c) Eclosing rate of pupa and lifespan of elav\u003e\u003eCG3814-RNAi. d)\nCrawling ability of larvae, climbing ability and lifespan of adults of Mef2\u003e\u003eCG3814-RNAi. e)\nActivity of Pdf\u003e\u003eCG3814-RNAi flies under 12/12 hours light/dark cycle. Data are represented\nas mean ± SD. One-way ANOVA followed by Newman-Keul’s multiple comparison test for\nmultiple comparison and two-way ANOVA followed by Sidak’s multiple comparison test for multiple comparison under two independent conditions were performed using GraphPad\nPrism. *p\u003c0.05.","description":"","filename":"Fig6.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/e13dddd3f4ab16f687256fc8.JPG"},{"id":3985226,"identity":"b36fe4fe-f441-417e-b056-7714c018597a","added_by":"auto","created_at":"2020-12-03 15:10:13","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":165767,"visible":true,"origin":"","legend":"Expression levels and phenotype of CG9722 (putative dmTMBIM1) knockdown\nin flies a) Temporal and tissue expression of CG9722/dmTMBIM1 from Flybase (http://flybase.org/).\nb) Viability of CG9722 knockdown (KD) flies by different GAL4 drivers and the KD efficiency\nof UAS- CG9722-RNAi (#1) assessed by quantitative PCR in whole larvae of Mef2\u003e\u003eCG9722- 18 RNAi. c) Hatching rate of pupa, adults’ wing phenotype and lifespan of elav\u003e\u003eCG9722-RNAi. d) Crawling ability of larvae, lifespan of a few surviving adults of Mef2\u003e\u003eCG9722-RNAi. e)\nActivity of Pdf\u003e\u003eCG9722-RNAi flies under 12/12 hours light/dark cycle. f)g) The transcriptional\nlevel of CG7188 and CG9722 in Mef\u003e\u003eCG9722-RNAi and Mef\u003e\u003eCG7188-RNAi respectively.\nData are represented as mean ± SD. One-way ANOVA followed by Newman-Keul’s multiple\ncomparison test for multiple comparison and unpaired t-test for two groups comparison were\nperformed using GraphPad Prism. *p\u003c0.05.","description":"","filename":"Fig7.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/e4f3e206f34810ce11d215fa.JPG"},{"id":3985227,"identity":"47f609de-c831-43ed-b775-dde2fa9ea750","added_by":"auto","created_at":"2020-12-03 15:10:13","extension":"jpg","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":282747,"visible":true,"origin":"","legend":"ER calcium and ER stress in TMBIM KD flies and summary of the phenotypes\na) Exemplary confocal images of ER calcium (erGAP3) in Pdf-GAL4 driven TMBIM knockdown\n(KD) flies and b) Quantification. Scale bar, 5 μm. c) Relative location of primers/probe used to\ndetect spliced and unspliced Xbp1 and d) Quantification of ER stress using the ratio of spliced\nand unspliced Xbp1 in heads from elav-GAL4 driven TMBIM KD flies. Each data point\nrepresents one animal in b) and one group (10 fly heads per group) in d). Data are represented\nas mean ± SD. One-way ANOVA followed by Newman-Keul’s multiple comparison test was\nperformed using GraphPad Prism. *p\u003c0.05. e) Summary of phenotypes observed in this study.","description":"","filename":"Fig8.JPG","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/1fc7f73e688a944b61ebd550.JPG"},{"id":13561701,"identity":"24f83bcd-7317-42bf-a661-b90303b833cf","added_by":"auto","created_at":"2021-09-17 03:10:07","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2517300,"visible":true,"origin":"","legend":"","description":"","filename":"LiTMBIMDM16.pdf","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1_covered.pdf"},{"id":13295770,"identity":"4d374941-cd80-4a23-a290-7bce03dc3fad","added_by":"auto","created_at":"2021-09-12 16:49:55","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2513735,"visible":true,"origin":"","legend":"","description":"","filename":"LiTMBIMDM16.pdf","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1_covered.pdf"},{"id":3985229,"identity":"58a40d63-7ac9-4cd1-acec-cef46e4d04f3","added_by":"auto","created_at":"2020-12-03 15:10:20","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1676961,"visible":true,"origin":"","legend":"","description":"","filename":"LiTMBIMDM16.pdf","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1_stamped.pdf"},{"id":3985220,"identity":"ee65f99e-a99c-4ccf-8657-9fbec07fc991","added_by":"auto","created_at":"2020-12-03 15:10:12","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":4100579,"visible":true,"origin":"","legend":"Supplemental Figure S1 Expression levels of CG9722 (dmTMBIM1) in the CG9722 RNAi#2 line and MiMIC line a) tub\u003e\u003eCG9722-RNAi#2 (BL6679) showed increased transcriptional levels of CG9722 assessed by quantitative qPCR in whole adults. b) CG9722-MiMIC line (BL44979), which is supposed to be a CG9722 knockout does not alter transcriptional levels of CG9722. c) Location of primers in respect to the CG9722 MiMIC insertion cassette. In homozygous CG9722 MiMIC flies, PCR using primer pair 1 amplifies a product of over 500 bp indicating no insertion of the MiMIC cassette. Primer pair 2 suggests the existence of the MiMIC cassette in the genome. Primer pairs 1for/2rev and 2for/1rev respectively 1for/2for and 1rev/2rev did not result in any PCR product in line with the lack of the MIMIC cassette within the CG9722 locus. Data are represented as mean ± SD. Unpaired t-test were performed using GraphPad Prism. *p\u003c0.05.\nSupplemental Table S1 RNAi lines used in this study.","description":"","filename":"SupplementaryInformation.docx","url":"https://assets-eu.researchsquare.com/files/rs-113374/v1/1ea08c590c3d0c346ff8a31b.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003eThe Evolutionary Conserved Transmembrane BAX Inhibitor Motif (TMBIM) Containing Protein Family is Essential for the Development and Survival of \u003cem\u003eDrosophila Melanogaster\u003c/em\u003e\u003c/p\u003e","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-113374/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"RECS1, PP1201, LFG3, MST100, MSTP100, NMP35, LFG, FAIM2, LFG2, NGP35, KIAA0950, LFG1, HNRGW, NMDARA1, MGC99687, GRINA, GBP, GAAP, S1R, LFG4, ZPRO, CGI-119, GHITM, MICS1, DERP2, My021, PTD010, HSPC282, FLJ26584, DKFZp566C0746","lastPublishedDoi":"10.21203/rs.3.rs-113374/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-113374/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"The Transmembrane BAX Inhibitor Motif (TMBIM) protein family consists of six evolutionarily conserved hydrophobic proteins that affect programmed cell death and the regulation of intracellular calcium levels. The bacterial orthologue BsYetJ is a pH-dependent calcium channel. We here identified six TMBIM family members in Drosophila melanogaster and studied the effect of their RNAi-mediated knockdown using ubiquitous and tissue-specific drivers. Mammalian TMBIM6 and TMBIM5 have obvious orthologs while this is more dubious for the other family members. Ubiquitous knockdown of family members dmTMBIM1,4,5, and 6 caused failed eclosing and tissue-specific knockdown resulted in a dramatically decreased lifespan. On the contrary, knockdown of dmTMBIM3, surprisingly, extended lifespan. Only knockdown of dmTMBIM6 increased the ER calcium levels of Pdf neurons. Neural knockdown of dmTMBIM2,3, and 4 increased ER stress, as indicated by increased Xbp1 splicing. Interestingly, TMBIM1 and TMBIM6 have a very similar expression pattern and their knockdown phenocopied each other. Also, knockdown of TMBIM1 resulted in upregulation of TMBIM6 and vice versa further suggesting a genetic interaction between these two genes. Our data demonstrate that most TMBIM proteins are essential for fly development and survival but, despite their shared protein structure, affect cell survival through different mechanisms.","manuscriptTitle":"The Evolutionary Conserved Transmembrane BAX Inhibitor Motif (TMBIM) Containing Protein Family is Essential for the Development and Survival of Drosophila Melanogaster","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-12-03 15:10:10","doi":"10.21203/rs.3.rs-113374/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"378d1d40-9abf-4a65-a64c-e49e14e03eb5","owner":[],"postedDate":"December 3rd, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":1319586,"name":"Cell Survival and Cell Death"},{"id":1319587,"name":"General Cell Biology \u0026 Physiology"}],"tags":[],"updatedAt":"2021-09-12T16:49:42+00:00","versionOfRecord":{"articleIdentity":"rs-113374","link":"https://doi.org/10.3389/fcell.2021.666484","journal":{"identity":"frontiers-in-cell-and-developmental-biology","isVorOnly":true,"title":"Frontiers in Cell and Developmental Biology"},"publishedOn":"2021-09-03 00:00:00","publishedOnDateReadable":"September 3rd, 2021"},"versionCreatedAt":"2020-12-03 15:10:10","video":"","vorDoi":"10.3389/fcell.2021.666484","vorDoiUrl":"https://doi.org/10.3389/fcell.2021.666484","workflowStages":[]},"version":"v1","identity":"rs-113374","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-113374","identity":"rs-113374","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}