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      Testis-specific ATP synthase peripheral stalk subunits required for tissue-specific mitochondrial morphogenesis in Drosophila

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          Abstract

          Background

          In Drosophila early post-meiotic spermatids, mitochondria undergo dramatic shaping into the Nebenkern, a spherical body with complex internal structure that contains two interwrapped giant mitochondrial derivatives. The purpose of this study was to elucidate genetic and molecular mechanisms underlying the shaping of this structure.

          Results

          The knotted onions ( knon) gene encodes an unconventionally large testis-specific paralog of ATP synthase subunit d and is required for internal structure of the Nebenkern as well as its subsequent disassembly and elongation. Knon localizes to spermatid mitochondria and, when exogenously expressed in flight muscle, alters the ratio of ATP synthase complex dimers to monomers. By RNAi knockdown we uncovered mitochondrial shaping roles for other testis-expressed ATP synthase subunits.

          Conclusions

          We demonstrate the first known instance of a tissue-specific ATP synthase subunit affecting tissue-specific mitochondrial morphogenesis. Since ATP synthase dimerization is known to affect the degree of inner mitochondrial membrane curvature in other systems, the effect of Knon and other testis-specific paralogs of ATP synthase subunits may be to mediate differential membrane curvature within the Nebenkern.

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          Most cited references52

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          Identification of functional elements and regulatory circuits by Drosophila modENCODE.

          To gain insight into how genomic information is translated into cellular and developmental programs, the Drosophila model organism Encyclopedia of DNA Elements (modENCODE) project is comprehensively mapping transcripts, histone modifications, chromosomal proteins, transcription factors, replication proteins and intermediates, and nucleosome properties across a developmental time course and in multiple cell lines. We have generated more than 700 data sets and discovered protein-coding, noncoding, RNA regulatory, replication, and chromatin elements, more than tripling the annotated portion of the Drosophila genome. Correlated activity patterns of these elements reveal a functional regulatory network, which predicts putative new functions for genes, reveals stage- and tissue-specific regulators, and enables gene-expression prediction. Our results provide a foundation for directed experimental and computational studies in Drosophila and related species and also a model for systematic data integration toward comprehensive genomic and functional annotation.
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            Mitochondrial dynamics and inheritance during cell division, development and disease.

            During cell division, it is critical to properly partition functional sets of organelles to each daughter cell. The partitioning of mitochondria shares some common features with that of other organelles, particularly in the use of interactions with cytoskeletal elements to facilitate delivery to the daughter cells. However, mitochondria have unique features - including their own genome and a maternal mode of germline transmission - that place additional demands on this process. Consequently, mechanisms have evolved to regulate mitochondrial segregation during cell division, oogenesis, fertilization and tissue development, as well as to ensure the integrity of these organelles and their DNA, including fusion-fission dynamics, organelle transport, mitophagy and genetic selection of functional genomes. Defects in these processes can lead to cell and tissue pathologies.
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              FACS-optimized mutants of the green fluorescent protein (GFP).

              We have constructed a library in Escherichia coli of mutant gfp genes (encoding green fluorescent protein, GFP) expressed from a tightly regulated inducible promoter. We introduced random amino acid (aa) substitutions in the twenty aa flanking the chromophore Ser-Tyr-Gly sequence at aa 65-67. We then used fluorescence-activated cell sorting (FACS) to select variants of GFP that fluoresce between 20-and 35-fold more intensely than wild type (wt), when excited at 488 nm. Sequence analysis reveals three classes of aa substitutions in GFP. All three classes of mutant proteins have highly shifted excitation maxima. In addition, when produced in E. coli, the folding of the mutant proteins is more efficient than folding of wt GFP. These two properties contribute to a greatly increased (100-fold) fluorescence intensity, making the mutants useful for a number of applications.
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                Author and article information

                Contributors
                sawyer.ericm@gmail.com
                lizbrunner18@gmail.com
                yihharn@gmail.com
                laivey89@gmail.com
                olivia.brown2019@gmail.com
                megbann@gmail.com
                deakrobetu@davidson.edu
                kesheaffer@gmail.com
                oshaunamorgan@gmail.com
                cofield@udel.edu
                nishita.suresh@gmail.com
                m.gracie.gordon@gmail.com
                tagunnell@hotmail.com
                lindsay056@gmail.com
                c_wood@lifesci.ucsb.edu
                mtfuller@stanford.edu
                kahales@davidson.edu
                Journal
                BMC Cell Biol
                BMC Cell Biol
                BMC Cell Biology
                BioMed Central (London )
                1471-2121
                23 March 2017
                23 March 2017
                2017
                : 18
                : 16
                Affiliations
                [1 ]ISNI 0000 0001 0531 1535, GRID grid.254902.8, Department of Biology, , Davidson College, ; Davidson, NC USA
                [2 ]ISNI 0000000419368956, GRID grid.168010.e, Departments of Developmental Biology and Genetics, , Stanford University School of Medicine, ; Stanford, CA USA
                Article
                132
                10.1186/s12860-017-0132-1
                5364652
                28335714
                2ac74f66-4323-4527-84fd-f9afce306b27
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 18 November 2016
                : 15 March 2017
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/100000001, National Science Foundation;
                Award ID: RUI 1158024
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000011, Howard Hughes Medical Institute;
                Award ID: Predoctoral Fellowship
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100000002, National Institutes of Health;
                Award ID: 1RO1-HD29194
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2017

                Cell biology
                drosophila melanogaster,spermatogenesis,mitochondria,atp synthase,cristae
                Cell biology
                drosophila melanogaster, spermatogenesis, mitochondria, atp synthase, cristae

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