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Suppression strategies utilizing genetically modified disease-refractory insects have surfaced as an attractive means of disease control, and progress has been made on engineering disease-resistant insect vectors. However, laboratory-engineered disease refractory genes would probably not spread in the wild, and would most likely need to be linked to a gene drive system in order to proliferate in native insect populations. Underdominant systems like translocations and engineered underdominance have been proposed as potential mechanisms for spreading disease refractory genes. Not only do these threshold-dependent systems have certain advantages over other potential gene drive mechanisms, such as localization of gene drive and removability, extreme engineered underdominance can also be used to bring about reproductive isolation, which may be of interest in controlling the spread of GMO crops. Proof-of-principle establishment of such drive mechanisms in a well-understood and studied insect, such as Drosophila melanogaster, is essential before more applied systems can be developed for the less characterized vector species of interest, such as mosquitoes. This work details the development of several distinct types of engineered underdominance and of translocations in Drosophila, including ones capable of bringing about reproductive isolation and population replacement, as a proof of concept study that can inform efforts to construct such systems in insect disease vectors.", "date": "2014", "date_type": "degree", "id_number": "CaltechTHESIS:05262014-104457839", "refereed": "FALSE", "official_url": "https://resolver.caltech.edu/CaltechTHESIS:05262014-104457839", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "collection": "CaltechTHESIS", "reviewer": "Kathy Johnson", "suggestions": "Released 09222015. Withheld until 1 July 2015. //\r\n\r\nPostdoc: abuchman@caltech.edu\r\n", "deposited_by": "Anna Basalova Buchman", "deposited_on": "2015-09-22 22:59:38", "doi": "10.7907/3YMP-DB79", "divisions": { "items": [ "div_bbe" ] }, "institution": "California Institute of Technology", "thesis_type": "phd", "thesis_advisor": { "items": [ { "email": "haybruce@caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "orcid": "0000-0002-5486-0482", "role": "advisor" } ] }, "thesis_committee": { "items": [ { "email": "zinnk@caltech.edu", "id": "Zinn-K-G", "name": { "family": "Zinn", "given": "Kai George" }, "orcid": "0000-0002-6706-5605", "role": "chair" }, { "email": "pws@caltech.edu", "id": "Sternberg-P-W", "name": { "family": "Sternberg", "given": "Paul W." }, "orcid": "0000-0002-7699-0173", "role": "member" }, { "email": "aravin@caltech.edu", "id": "Aravin-A-A", "name": { "family": "Aravin", "given": "Alexei A." }, "orcid": "0000-0002-6956-8257", "role": "member" }, { "email": "haybruce@caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "orcid": "0000-0002-5486-0482", "role": "member" } ] }, "thesis_degree": "PHD", "thesis_degree_grantor": "California Institute of Technology", "thesis_defense_date": "2014-05-28", "gradofc_approval_date": "2014-06-02", "thesis_awards": "Lawrence L. and Audrey W. Ferguson Prize, 2014\r\n", "review_status": "approved", "option_major": { "items": [ "biology" ] }, "copyright_statement": "Author's Rights Authorization: I hereby certify that, if appropriate, I have obtained a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted here is the same as that approved by my advisory committee.\n\nI hereby grant to California Institute of Technology or its agents the non-exclusive license to archive and make accessible, under the conditions specified under \"Thesis Availability\" in this submission, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation, or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.", "resource_type": "thesis", "pub_year": "2014", "author_list": "Basalova Buchman, Anna", "advisor_list": "Hay, Bruce A.", "comittee_list": "Zinn, Kai George; Sternberg, Paul W.; et el." }, { "id": "https://thesis.library.caltech.edu/id/eprint/7055", "eprint_id": 7055, "rev_number": 30, "documents": [ { "id": "/id/document/44837", "doc_id": 44837, "rev_number": 3, "files": [ { "id": "/id/file/132564", "fileid": 132564, "datasetid": "document", "objectid": 44837, "filename": "CaltechTHESIS PhD thesis release request for Matzen Kelly Jean (2012).txt", "mime_type": "message/rfc822\\0117bit", "hash": "d3a05a77a2bcbfa8b263e601732f0627", "hash_type": "MD5", "filesize": 1487, "mtime": "2014-01-10 00:21:28", "url": "/7055/2/CaltechTHESIS PhD thesis release request for Matzen Kelly Jean (2012).txt" } ], "eprint_id": 7055, "pos": 2, "placement": 2, "mime_type": 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"media_sample_stop": "0", "relation": { "items": [ { "type": "http://eprints.org/relation/isVersionOf", "uri": "/id/document/45080" }, { "type": "http://eprints.org/relation/isVolatileVersionOf", "uri": "/id/document/45080" }, { "type": "http://eprints.org/relation/isIndexCodesVersionOf", "uri": "/id/document/45080" } ] } } ], "eprint_status": "archive", "userid": 4049, "dir": "disk0/00/00/70/55", "datestamp": "2014-01-29 18:17:42", "lastmod": "2019-10-03 23:55:59", "status_changed": "2014-01-29 18:17:42", "type": "thesis", "metadata_visibility": "show", "creators": { "items": [ { "email": "matzenkj@gmail.com", "id": "Matzen-Kelly-Jean", "name": { "family": "Matzen", "given": "Kelly Jean" }, "show_email": "YES" } ] }, "title": "Engineering of Dengue Virus Refractoriness in Aedes aegypti and Development of an Underdominant Gene Drive System in Drosophila melanogaster", "ispublished": "unpub", "full_text_status": "public", "keywords": "Gene drive; Underdominance; Drosophila; Dengue; Engineering; Synthetic Biology; Reproductive Isolation; Viral Refractoriness", "abstract": "
Vector-borne diseases have a profound impact on world health. The two most well-known and costly diseases are dengue fever and malaria, both spread by mosquito vectors. In the last decade, many new solutions to halting the spread of these diseases have been sought, including vector-mediated disease suppression. The work presented here seeks to generate alleles to effect this suppression, and engineer a drive system to replace the native population. Additional work on systems to keep engineered organisms genetically isolated from native populations has also been carried out. Initial studies in C. elegans investigated use of the transitive nature of RNAi in this species to genetically isolate one population from another. This type of speciation could be used in plant populations to limit gene flow of engineered crops into local environments.
\r\n\r\nThe next series of studies details work on engineering of refractoriness alleles. Dengue virus has several enzymatic activities that are essential for its replicative cycle, including an RNA-dependednt RNA polymerase (RdRp) responsible for synthesizing both the sense and antisense viral genomes and a protease responsible for several essential cleavages of the viral polyprotein. Artificial substrates for these proteins were created to act as sensors, triggering an apoptotic response when viral infection occurs. Several generations of constructs were tested, but so far no completely functional sensor has been generated.
\r\n\r\nLastly, a series of underdominant gene drive architectures were built and tested in Drosophila melanogaster. Initial systems utilized a Drosophila cell death protein, Hid, as toxin, and engineered microRNAs designed to target the Hid proteins as antidote. Two toxin-antidote pairs were mismatched and positioned on separate chromosomes so that an organism carrying both chromosomes survives, but an organism carrying only a single chromosome is unviable. Construction of a proof-of-principle in the eye was successful, but work in essential tissues is ongoing. Systems using engineered microRNAs as toxins and resupply of the native protein as antidote were tested in essential tissues. Testing of many components has contributed to the development of these systems, but a complete system has not yet been constructed.
", "date": "2012", "date_type": "degree", "id_number": "CaltechTHESIS:05192012-130917004", "refereed": "FALSE", "official_url": "https://resolver.caltech.edu/CaltechTHESIS:05192012-130917004", "projects": { "items": [ "Dengue Virus Refractoriness", "Underdominance", "Transitive RNAi Reproductive Isolation" ] }, "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "collection": "CaltechTHESIS", "reviewer": "Kathy Johnson", "deposited_by": "Kelly Matzen", "deposited_on": "2014-01-29 18:17:42", "doi": "10.7907/YNFQ-WM56", "divisions": { "items": [ "div_biol" ] }, "institution": "California Institute of Technology", "thesis_type": "phd", "thesis_advisor": { "items": [ { "email": "haybruce@caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "role": "advisor" } ] }, "thesis_committee": { "items": [ { "email": "straussj@its.caltech.edu", "id": "Strauss-J-H", "name": { "family": "Strauss", "given": "James H." }, "role": "chair" }, { "email": "bjorkman@caltech.edu", "id": "Bjorkman-P-J", "name": { "family": "Bjorkman", "given": "Pamela J." }, "role": "member" }, { "email": "baltimo@caltech.edu", "id": "Baltimore-D-L", "name": { "family": "Baltimore", "given": "David L." }, "role": "member" }, { "email": "haybruce@caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "role": "member" } ] }, "thesis_degree": "PHD", "thesis_degree_grantor": "California Institute of Technology", "thesis_defense_date": "2012-05-02", "gradofc_approval_date": "2012-05-22", "review_status": "approved", "option_major": { "items": [ "biochem" ] }, "copyright_statement": "Author's Rights Authorization: I hereby certify that, if appropriate, I have obtained a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted here is the same as that approved by my advisory committee.\n\nI hereby grant to California Institute of Technology or its agents the non-exclusive license to archive and make accessible, under the conditions specified under \"Thesis Availability\" in this submission, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation, or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.", "resource_type": "thesis", "pub_year": "2012", "author_list": "Matzen, Kelly Jean", "advisor_list": "Hay, Bruce A.", "comittee_list": "Strauss, James H.; Bjorkman, Pamela J.; et el." }, { "id": "https://thesis.library.caltech.edu/id/eprint/11763", "eprint_id": 11763, "rev_number": 11, "documents": [ { "id": "/id/document/102737", "doc_id": 102737, "rev_number": 2, "files": [ { "id": "/id/file/296089", "fileid": 296089, "datasetid": "document", "objectid": 102737, "filename": "CEWard-ThesisDone.pdf", "mime_type": "application/pdf", "hash": "10867b2e355c4cd4d320ce21ff55b6ef", "hash_type": "MD5", "filesize": 3810119, "mtime": "2019-08-20 20:24:57", "url": "/11763/1/CEWard-ThesisDone.pdf" } ], "eprint_id": 11763, "pos": 1, "placement": 1, "mime_type": "application/pdf", "format": "application/pdf", "language": "en", "security": "public", "license": 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element, Medea, mosquito, malaria", "abstract": "Insect-borne diseases kill millions of people annually. One strategy for controlling transmission of insect-borne disease involves replacing the native insect population with transgenic animals unable to transmit disease. Population replacement requires a drive mechanism to ensure the rapid spread of linked transgenes conferring disease refractoriness. Medea selfish genetic elements have the feature that when present in a female, only offspring that inherit the element survive, a behavior that can lead to spread. Here we use modeling to identify conditions under which Medea elements spread. We derive equations describing the allele frequencies required for spread of Medea elements with a fitness cost, and the equilibrium allele frequencies attained. We validate our model against a synthetic Medea element created in Drosophila and find that the model fits the data without parameter fitting. We show that when Medea spreads, it drives the non-Medea genotype out of the population, and we provide estimates of the number of generations required to achieve this goal. We also characterize two contexts in which Medea elements with fitness costs drive the non-Medea allele from the population: an autosomal element in which zygotic rescue is incomplete and an X-linked element in species in which X/Y individuals are male. Finally, we explore costs and benefits associated with the introduction of multiple Medea elements. Our results suggest that Medea elements can drive population replacement under a wide range of conditions, potentially reducing disease burden.", "date": "2011", "date_type": "degree", "id_number": "CaltechTHESIS:05312011-123357339", "refereed": "FALSE", "official_url": "https://resolver.caltech.edu/CaltechTHESIS:05312011-123357339", "referencetext": { "items": [ "Asman, S. M., P. T. McDonald, and T. Prout. 1981. 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Genet Res 68:125-129." ] }, "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "funders": { "items": [ { "agency": "NIH", "grant_number": "GM057422" }, { "agency": "NIH", "grant_number": "GM70956" }, { "agency": "NIH", "grant_number": "R01 GM072879" }, { "agency": "NIH", "grant_number": "R01 GM070956" }, { "agency": "NIH", "grant_number": "DP1 OD003878" }, { "agency": "Regents of the University of California from the Foundation for the National Institutes of Health through the Grand Challenges in Global Health initiative" } ] }, "collection": "CaltechTHESIS", "reviewer": "Kathy Johnson", "deposited_by": "Kathy Johnson", "deposited_on": "2019-08-20 20:26:23", "doi": "10.7907/T656-ZN91", "divisions": { "items": [ "div_biol" ] }, "institution": "California Institute of Technology", "thesis_type": "phd", "thesis_advisor": { "items": [ { "email": "haybruce@caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce" }, "role": "advisor" } ] }, "thesis_committee": { "items": [ { "email": "pws@caltech.edu", "id": "Sternberg-P-W", "name": { "family": "Sternberg", "given": "Paul W." }, "role": "chair" }, { "email": "melowitz@caltech.edu", "id": "Elowitz-M-B", "name": { "family": "Elowitz", "given": "Michael B." }, "role": "member" }, { "email": "murray@caltech.edu", "id": "Murray-R-M", "name": { "family": "Murray", "given": "Richard M." }, "role": "member" }, { "email": "haybruce@caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "role": "member" } ] }, "thesis_degree": "PHD", "thesis_degree_grantor": "California Institute of Technology", "thesis_defense_date": "2010-12-20", "gradofc_approval_date": "2011-05-31", "review_status": "approved", "option_major": { "items": [ "biochem" ] }, "copyright_statement": "Author's Rights Authorization: I hereby certify that, if appropriate, I have obtained a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted here is the same as that approved by my advisory committee.\n\nI hereby grant to California Institute of Technology or its agents the non-exclusive license to archive and make accessible, under the conditions specified under \"Thesis Availability\" in this submission, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation, or project report. 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"media_duration": "0", "media_aspect_ratio": "0", "media_sample_start": "0", "media_sample_stop": "0", "relation": { "items": [ { "type": "http://eprints.org/relation/isVersionOf", "uri": "/id/document/4481" }, { "type": "http://eprints.org/relation/isVolatileVersionOf", "uri": "/id/document/4481" }, { "type": "http://eprints.org/relation/isIndexCodesVersionOf", "uri": "/id/document/4481" } ] } }, { "id": "/id/document/122744", "doc_id": 122744, "rev_number": 1, "files": [ { "id": "/id/file/368142", "fileid": 368142, "datasetid": "document", "objectid": 122744, "filename": "indexcodes.txt", "mime_type": "text/plain", "hash": "45efd9fbdff91d3b21026ba6d15eedb6", "hash_type": "MD5", "filesize": 3063, "mtime": "2021-06-23 22:05:59", "url": "/2356/18/indexcodes.txt" } ], "eprint_id": 2356, "pos": 18, "placement": 18, "mime_type": "text/plain", "format": "other", "format_desc": "Generate index codes conversion from application/pdf to indexcodes", "language": "en", "security": "public", "license": "other", "main": "indexcodes.txt", "media_duration": "0", "media_aspect_ratio": "0", "media_sample_start": "0", "media_sample_stop": "0", "relation": { "items": [ { "type": "http://eprints.org/relation/isVersionOf", "uri": "/id/document/4482" }, { "type": "http://eprints.org/relation/isVolatileVersionOf", "uri": "/id/document/4482" }, { "type": "http://eprints.org/relation/isIndexCodesVersionOf", "uri": "/id/document/4482" } ] } } ], "eprint_status": "archive", "userid": 2, "dir": "disk0/00/00/23/56", "datestamp": "2005-06-01", "lastmod": "2020-05-08 21:25:50", "status_changed": "2009-09-25 02:35:34", "type": "thesis", "metadata_visibility": "show", "creators": { "items": [ { "email": "jeffrey.copeland@emu.edu", "id": "Copeland-Jeffrey-Michael", "name": { "family": "Copeland", "given": "Jeffrey Michael" }, "orcid": "0000-0001-5432-3524", "show_email": "NO" } ] }, "title": "Identification of Novel Cell Death Regulators in C. elegans and Drosophila", "ispublished": "unpub", "full_text_status": "public", "keywords": "apoptosis; Drosophila; echinus; eye", "abstract": "Apoptosis is a form of cell death executed by a class of cysteine proteases called caspases. Though caspases are well-conserved, the mechanisms by which caspases are regulated vary remarkably. This dissertation addresses three independent aspects of apoptosis and its regulation.
\r\n\r\nIn the developing Drosophila eye, apoptosis is activated to remove extra cells that are initially present between ommatidia. Mutants for the gene echinus have a disorganized eye structure due to a failure of these cell deaths to occur. We demonstrate that echinus resembles a deubiquitinating enzyme, that it is expressed in the pupal eye during the time of cell death, and that echinus acts genetically upstream or independently of the death-inducing genes head involution defective, reaper, and grim. Based on in vitro assays and the fact that the Echinus enzyme lacks a catalytic cysteine residue, we propose that echinus and its orthologs constitute a novel class of inactive deubiquitinating enzymes, perhaps functioning in a dominant-negative manner to inhibit deubiquitination of specific substrates.
\r\n\r\nIn C. elegans, the model for caspase inhibition is quite different from that in Drosophila and in mammals. To look for genes that directly inhibit the CED-3 caspase, we screened a C. elegans cDNA library for CED-3 suppressors in the yeast S. cerevisiae and found several suppressors. Experiments in yeast suggest that one of these genes, Y39B6A.12, requires the prodomain of CED-3 for suppression, and ectopic expression in the Drosophila eye shows that it can suppress apoptosis induced by the Bcl2 family member Debcl.
\r\n\r\nIn Drosophila, DIAP1 is the focal point in the regulation of apoptosis. To identify novel regulators of DIAP1, deficiency chromosomes spanning the Drosophila genome were screened for dominant modifiers of a diap1 knockdown phenotype. Nine deficiencies were isolated that cover no known regulators, and two modifiers were mapped to small genomic regions. This screen has provided a starting point for identifying some of the many uncharacterized genes that are involved in regulating apoptosis.
", "date": "2005", "date_type": "degree", "id_number": "CaltechETD:etd-06012005-114127", "refereed": "FALSE", "official_url": "https://resolver.caltech.edu/CaltechETD:etd-06012005-114127", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "collection": "CaltechTHESIS", "reviewer": "Kathy Johnson", "deposited_by": "Imported from ETD-db", "deposited_on": "2005-06-01", "doi": "10.7907/7T3M-DY09", "divisions": { "items": [ "div_biol" ] }, "institution": "California Institute of Technology", "thesis_type": "phd", "thesis_advisor": { "items": [ { "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "role": "advisor" } ] }, "thesis_committee": { "items": [ { "email": "deshaies@its.caltech.edu", "id": "Deshaies-R-J", "name": { "family": "Deshaies", "given": "Raymond Joseph" }, "role": "chair" }, { "email": "haybruce@its.caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "role": "member" }, { "email": "meyerow@its.caltech.edu", "id": "Meyerowitz-E-M", "name": { "family": "Meyerowitz", "given": "Elliot M." }, "role": "member" }, { "email": "zinnk@its.caltech.edu", "id": "Zinn-K-G", "name": { "family": "Zinn", "given": "Kai George" }, "role": "member" }, { "email": "pws@its.caltech.edu", "id": "Sternberg-P-W", "name": { "family": "Sternberg", "given": "Paul W." }, "role": "member" } ] }, "thesis_degree": "PHD", "thesis_degree_grantor": "California Institute of Technology", "thesis_submitted_date": "2005-06-01", "thesis_defense_date": "2005-05-11", "thesis_approved_date": "2005-06-01", "review_status": "approved", "option_major": { "items": [ "biology" ] }, "copyright_statement": "I hereby certify that, if appropriate, I have obtained a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee.\n\nI hereby grant to California Institute of Technology or its agents the non-exclusive\nlicense to archive and make accessible, under the conditions specified below,\nmy thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.", "resource_type": "thesis", "pub_year": "2005", "author_list": "Copeland, Jeffrey Michael", "advisor_list": "Hay, Bruce A.", "comittee_list": "Deshaies, Raymond Joseph; Hay, Bruce A.; et el." }, { "id": "https://thesis.library.caltech.edu/id/eprint/2425", "eprint_id": 2425, "rev_number": 12, "documents": [ { "id": "/id/document/4607", "doc_id": 4607, "rev_number": 3, "files": [ { "id": "/id/file/26563", "fileid": 26563, "datasetid": "document", "objectid": 4607, "filename": "Jun_Huh_thesis.pdf", "mime_type": "application/pdf", "filesize": 5524400, "mtime": "2012-12-26 02:51:15", "url": "/2425/1/Jun_Huh_thesis.pdf" } ], "eprint_id": 2425, "pos": 1, "mime_type": "application/pdf", "format": "application/pdf", "format_desc": "Jun_Huh_thesis.pdf", "language": "en", "security": "public", "license": "other", "main": "Jun_Huh_thesis.pdf", "media_duration": 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"media_sample_stop": "0", "relation": { "items": [ { "type": "http://eprints.org/relation/isVersionOf", "uri": "/id/document/44184" }, { "type": "http://eprints.org/relation/isVolatileVersionOf", "uri": "/id/document/44184" }, { "type": "http://eprints.org/relation/isIndexCodesVersionOf", "uri": "/id/document/44184" } ] } } ], "eprint_status": "archive", "userid": 2, "dir": "disk0/00/00/24/25", "datestamp": "2005-06-03", "lastmod": "2020-05-20 21:10:14", "status_changed": "2009-09-25 02:37:17", "type": "thesis", "metadata_visibility": "show", "creators": { "items": [ { "email": "jun_huh@hms.harvard.edu", "id": "Huh-Jun-Ryul", "name": { "family": "Huh", "given": "Jun Ryul" }, "show_email": "YES" } ] }, "title": "To Die or to Differentiate: Apoptotic and Non-Apoptotic Roles of Death Molecules in Drosophila melanogaster", "ispublished": "unpub", "full_text_status": "public", "keywords": "apoptosis; caspases; diap; grim; hid; iap; programmed cell death; proliferation; rpr; spermatogenesis; stress; translational inhibition; ubiquitination", "abstract": "Virtually every cell, if not all, are ready to die under stressful conditions or by necessity during animal development. In Drosophila, three pro-apoptotic proteins, Rpr/Hid/Grim, had been found to induce cell death by preventing the function of the cell death inhibitor, DIAP1. However, the mechanistic details of this process were largely unknown. We have found that Rpr/Hid/Grim induce DIAP1 destabilization through ubiquitination and general translational inhibition. Moreover, from the in-vitro and in-vivo studies, we also found that ubiquitination of DIAP1 by Hid is dependant on DIAP1\u2019s own ability to ubiquitinate itself. Once the life-or-death decision is made, cells can efficiently start apoptosis by quickly removing pre-existing death inhibitors using these mechanisms.
\r\n\r\nIn addition to the canonical roles of death machinery, we have also studied their roles in non-apoptotic developmental processes. In the testis, germline stem cells ultimately give rise to 64 individual sperms. Spermatocytes, and later, spermatids, develop within a single membranous structure, or syncytium. Formation of free-swimming sperms requires the encapsulation of each spermatid by an independent plasma membrane and the elimination of most of the sperm cytoplasm. We demonstrated that at least three independent caspase activation pathways are likely to be involved in these processes with different spatial and temporal activation patterns, and that a global inhibition of caspase activity results in male sterility.
\r\n\r\nExternal stresses such as radiation and heat shock were known to induce large amounts of cell death (up to 60% of the total cell population) in proliferative tissues like Drosophila larval imaginal discs. Interestingly, larvae exposed to such stress ultimately develop into normal adult flies. This is facilitated by the compensatory proliferation of cells that neighbor the dying cells. In order to study the mechanistic basis for this process, we uncoupled cell death from death activation by expressing Hid in the presence of P35, a viral inhibitor of effecter caspases. Interestingly, neighboring cells of clones expressing Hid underwent compensatory proliferation, which was no longer observed when we blocked the activation of initiator caspase, Dronc. Our observations indicate that non-apoptotic Dronc activity is required for the generation of a non-autonomous proliferation signal.
", "date": "2005", "date_type": "degree", "id_number": "CaltechETD:etd-06032005-145438", "refereed": "FALSE", "official_url": "https://resolver.caltech.edu/CaltechETD:etd-06032005-145438", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "collection": "CaltechTHESIS", "reviewer": "Kathy Johnson", "deposited_by": "Imported from ETD-db", "deposited_on": "2005-06-03", "doi": "10.7907/Q00G-B953", "divisions": { "items": [ "div_biol" ] }, "institution": "California Institute of Technology", "thesis_type": "phd", "thesis_advisor": { "items": [ { "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "role": "advisor" } ] }, "thesis_committee": { "items": [ { "email": "zinnk@caltech.edu", "id": "Zinn-K-G", "name": { "family": "Zinn", "given": "Kai George" }, "role": "chair" }, { "email": "haybruce@caltech.edu", "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "role": "member" }, { "email": "wuwei@its.caltech.edu", "id": "Anderson-D-J", "name": { "family": "Anderson", "given": "David J." }, "role": "member" }, { "email": "pws@caltech.edu", "id": "Sternberg-P-W", "name": { "family": "Sternberg", "given": "Paul W." }, "role": "member" }, { "email": "dunphy@cco.caltech.edu", "id": "Dunphy-W-G", "name": { "family": "Dunphy", "given": "William G." }, "role": "member" } ] }, "thesis_degree": "PHD", "thesis_degree_grantor": "California Institute of Technology", "thesis_submitted_date": "2005-06-03", "thesis_defense_date": "2005-05-17", "thesis_approved_date": "2005-06-03", "review_status": "approved", "option_major": { "items": [ "biology" ] }, "copyright_statement": "I hereby certify that, if appropriate, I have obtained a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee.\n\nI hereby grant to California Institute of Technology or its agents the non-exclusive\nlicense to archive and make accessible, under the conditions specified below,\nmy thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.", "resource_type": "thesis", "pub_year": "2005", "author_list": "Huh, Jun Ryul", "advisor_list": "Hay, Bruce A.", "comittee_list": "Zinn, Kai George; Hay, Bruce A.; et el." }, { "id": "https://thesis.library.caltech.edu/id/eprint/6801", "eprint_id": 6801, "rev_number": 19, "documents": [ { "id": "/id/document/14166", "doc_id": 14166, "rev_number": 5, "files": [ { "id": "/id/file/91197", "fileid": 91197, "datasetid": "document", "objectid": 14166, "filename": "Vernooy_sy_2002.pdf", "mime_type": "application/pdf", "filesize": 27803339, "mtime": "2012-12-26 04:40:06", "url": "/6801/1/Vernooy_sy_2002.pdf" } ], "eprint_id": 6801, "pos": 1, "mime_type": "application/pdf", "format": "application/pdf", "language": "en", "security": "public", "license": "other", "main": "Vernooy_sy_2002.pdf", "media_duration": "0", "media_aspect_ratio": "0", "media_sample_start": "0", "media_sample_stop": "0", "content": "final", "relation": { "items": [ { "type": "http://eprints.org/relation/hasVolatileVersion", "uri": "/id/document/27787" }, { "type": "http://eprints.org/relation/haspreviewThumbnailVersion", "uri": "/id/document/27787" }, { "type": "http://eprints.org/relation/hasVersion", "uri": "/id/document/27787" } ] } }, { "id": "/id/document/14167", "doc_id": 14167, "rev_number": 4, "files": [ { "id": "/id/file/207712", "fileid": 207712, "datasetid": "document", "objectid": 14167, "filename": "Vernooy_sy_2002.zip", "mime_type": "application/zip", "filesize": 89363259, "mtime": "2016-08-22 21:23:33", "url": "/6801/2/Vernooy_sy_2002.zip" } ], "eprint_id": 6801, "pos": 2, "format": "application/zip", "format_desc": "TIFF", "language": "en", "security": "internal", "license": "other", "main": "Vernooy_sy_2002.zip", "media_duration": "0", "media_aspect_ratio": "0", "media_sample_start": "0", "media_sample_stop": "0", "content": "archival" }, { "id": "/id/document/27787", "doc_id": 27787, "rev_number": 3, "files": [ { "id": "/id/file/91195", "fileid": 91195, "datasetid": "document", "objectid": 27787, "filename": "preview.png", "mime_type": "image/png", "hash": "10daffbfb8ea32aab19cd92455ba0971", "hash_type": "MD5", "filesize": 40881, "mtime": "2012-12-26 04:40:06", "url": "/6801/3/preview.png" } ], "eprint_id": 6801, "pos": 3, "placement": 3, "mime_type": "image/png", "format": "image/png", "language": "en", "security": "public", "license": "other", "main": "preview.png", "media_duration": "0", "media_aspect_ratio": "0", "media_sample_start": "0", "media_sample_stop": "0", "relation": { "items": [ { "type": "http://eprints.org/relation/isVolatileVersionOf", "uri": "/id/document/14166" }, { "type": "http://eprints.org/relation/ispreviewThumbnailVersionOf", "uri": "/id/document/14166" }, { "type": "http://eprints.org/relation/isVersionOf", "uri": "/id/document/14166" } ] } }, { "id": "/id/document/127182", "doc_id": 127182, "rev_number": 1, "files": [ { "id": "/id/file/372616", "fileid": 372616, "datasetid": "document", "objectid": 127182, "filename": "indexcodes.txt", "mime_type": "text/plain", "hash": "808f3a010d243df6309d803ced70a90e", "hash_type": "MD5", "filesize": 24610, "mtime": "2021-06-24 01:31:29", "url": "/6801/4/indexcodes.txt" } ], "eprint_id": 6801, "pos": 4, "placement": 4, "mime_type": "text/plain", "format": "other", "format_desc": "Generate index codes conversion from application/pdf to indexcodes", "language": "en", "security": "public", "license": "other", "main": "indexcodes.txt", "media_duration": "0", "media_aspect_ratio": "0", "media_sample_start": "0", "media_sample_stop": "0", "relation": { "items": [ { "type": "http://eprints.org/relation/isVersionOf", "uri": "/id/document/14166" }, { "type": "http://eprints.org/relation/isVolatileVersionOf", "uri": "/id/document/14166" }, { "type": "http://eprints.org/relation/isIndexCodesVersionOf", "uri": "/id/document/14166" } ] } } ], "eprint_status": "archive", "userid": 87, "dir": "disk0/00/00/68/01", "datestamp": "2012-02-01 21:27:03", "lastmod": "2021-11-05 19:25:15", "status_changed": "2012-02-01 21:27:02", "type": "thesis", "metadata_visibility": "show", "creators": { "items": [ { "id": "Vernooy-Stephanie-Yeager", "name": { "family": "Vernooy", "given": "Stephanie Yeager" }, "show_email": "NO" } ] }, "title": "Identification of Apoptotic Regulators in Drosophila and their Nonapoptotic Roles in Spermatogenesis: Implications for the Existence of a \"Caspase Cassette\" which Regulates Diverse Biological Processes", "ispublished": "unpub", "full_text_status": "public", "keywords": "Biology", "abstract": "Drosophila has long been an attractive, genetically tractable model system in which to study fundamental processes such as apoptosis which are common to higher eukaryotes. Following completion of the Drosophila genome sequence, we carried out comprehensive BLAST searches to annotate it with respect to apoptosis, and found sequence homologues of virtually all mammalian cell death genes with the exception of death receptors. The only Drosophila cell death genes for which mammalian homologues have not been identified are the cell death activators Rpr, Hid, and Grim. However, since proteins with similar activities are present in mammals and since their mechanisms are likely to be conserved even if true sequence homologues are not identified, understanding how Rpr, Hid, and Grim act to bring about death is an important area of research. To better understand their mechanisms of action, we carried out an overexpression screen to identify suppressors of Rpr-, Hid-, and Grim-induced death. We identified the strongest of these suppressors as dBruce, a large protein with an N-terminal baculovirus IAP repeat (BIR), characteristic of inhibitors of apoptosis (IAPs), and a C-terminal ubiquitin conjugation domain (E2). We show that it potently suppresses death induced by Rpr and Grim but not by Hid, and that this activity likely requires its E2 domain. It does not directly promote degradation of Rpr or Grim, but its antiapoptotic action requires that their N-termini, through which they interact with BIR2 of DIAP1, be intact. These data, combined with the inability of dBruce to block death induced by the apical caspase Dronc or the proapoptotic Bcl-2 family member Debcl/Drob-1/dBorg-1/Dbok, suggest that dBruce regulates cell death at a novel point. Interestingly, dBruce mutant males are sterile, but a lack of increased caspase activity in these mutants suggests that dBruce may also play nonapoptotic roles. A closer look at Drosophila male testes revealed the surprising observation that high levels of caspases are present in wild type testes, along with the caspase activator Ark. This provokes speculation that core components of the cell death machinery can function to regulate processes other than apoptosis, such as spermatogenesis.", "date": "2002", "date_type": "degree", "id_number": "CaltechTHESIS:02012012-113408899", "refereed": "FALSE", "official_url": "https://resolver.caltech.edu/CaltechTHESIS:02012012-113408899", "rights": "No commercial reproduction, distribution, display or performance rights in this work are provided.", "collection": "CaltechTHESIS", "reviewer": "Tony Diaz", "deposited_by": "Benjamin Perez", "deposited_on": "2012-02-01 21:27:03", "doi": "10.7907/zqpk-k618", "divisions": { "items": [ "div_biol" ] }, "institution": "California Institute of Technology", "thesis_type": "phd", "thesis_advisor": { "items": [ { "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "orcid": "0000-0002-5486-0482", "role": "advisor" }, { "id": "Sternberg-P-W", "name": { "family": "Sternberg", "given": "Paul W." }, "orcid": "0000-0002-7699-0173", "role": "co-advisor" } ] }, "thesis_committee": { "items": [ { "id": "Bronner-M-E", "name": { "family": "Bronner", "given": "Marianne E." }, "orcid": "0000-0003-4274-1862", "role": "chair" }, { "id": "Sternberg-P-W", "name": { "family": "Sternberg", "given": "Paul W." }, "orcid": "0000-0002-7699-0173", "role": "co-chair" }, { "id": "Hay-B-A", "name": { "family": "Hay", "given": "Bruce A." }, "orcid": "0000-0002-5486-0482", "role": "member" }, { "id": "Deshaies-R-J", "name": { "family": "Deshaies", "given": "Raymond Joseph" }, "orcid": "0000-0002-3671-9354", "role": "member" }, { "id": "Zinn-K-G", "name": { "family": "Zinn", "given": "Kai George" }, "orcid": "0000-0002-6706-5605", "role": "member" } ] }, "thesis_degree": "PHD", "thesis_degree_grantor": "California Institute of Technology", "thesis_defense_date": "2002-05-29", "review_status": "approved", "option_major": { "items": [ "biology" ] }, "copyright_statement": "Author's Rights Authorization: I hereby certify that, if appropriate, I have obtained a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted here is the same as that approved by my advisory committee.\n\nI hereby grant to California Institute of Technology or its agents the non-exclusive license to archive and make accessible, under the conditions specified under \"Thesis Availability\" in this submission, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation, or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report.", "resource_type": "thesis", "pub_year": "2002", "author_list": "Vernooy, Stephanie Yeager", "advisor_list": "Hay, Bruce A. and Sternberg, Paul W.", "comittee_list": "Bronner, Marianne E.; Sternberg, Paul W.; et el." } ]