期刊论文详细信息
BMC Research Notes
Mitochondrial fission and fusion in Dictyostelium discoideum: a search for proteins involved in membrane dynamics
Kari Naylor1  Gregory W Berbusse1  Brixey G Schimmel1 
[1] Biology Department, University of Central Arkansas, 180 Lewis Science Center, Conway, AR, 72035, USA
关键词: Dynamin;    CluA;    MidA;    Dictyostelium discoideum;    Mitochondrial morphology;    Fusion;    Fission;    Mitochondria;   
Others  :  1165686
DOI  :  10.1186/1756-0500-5-505
 received in 2012-07-22, accepted in 2012-09-06,  发布年份 2012
PDF
【 摘 要 】

Background

Mitochondrial morphology is maintained by two distinct membrane events -fission and fusion. Altering these conserved processes can disrupt mitochondrial morphology and distribution, thereby disrupting the organelle’s functionality and impeding cellular function. In higher eukaryotes, these processes are mediated by a family of dynamin-related proteins (DRP’s). In the lower eukaryotes, for instance Dictyostelium discoideum, mitochondrial fission and fusion have been implicated but not yet established. To understand the overall mechanism of these dynamics across organisms, we developed an assay to identify fission and fusion events in Dictyostelium and to assess the involvement of the mitochondrial proteins, MidA, CluA, and two DRP’s, DymA and DymB.

Findings

Using laser scanning confocal microscopy we show, for the first time, that lower eukaryotes mediate mitochondrial fission and fusion. In Dictyostelium, these processes are balanced, occurring approximately 1 event/minute. Quantification of the rates in midA-, cluA-, dymA-, or dymB- strains established that MidA appears to play an indirect role in the regulation of fission and fusion, while the DRP’s are not essential for these processes. Rates of fission and fusion were significantly reduced in cluA-cells, indicating that CluA is necessary for maintaining both fission and fusion.

Conclusions

We have successfully demonstrated that Dictyostelium mitochondria undergo the dynamic processes of fission and fusion. The classical mediators of membrane dynamics - the DRP’s – are not necessary for these dynamics, whereas CluA is necessary for both processes. This work contributes to our overall understanding of mitochondrial dynamics and ultimately will provide additional insight into mitochondrial disease.

【 授权许可】

   
2012 Schimmel et al.; licensee BioMed Central Ltd.

【 预 览 】
附件列表
Files Size Format View
20150416032824800.pdf 2753KB PDF download
Figure 5 . 116KB Image download
Figure 4 . 89KB Image download
Figure 3 . 75KB Image download
Figure 2 . 100KB Image download
Figure 1 . 99KB Image download
【 图 表 】

Figure 1 .

Figure 2 .

Figure 3 .

Figure 4 .

Figure 5 .

【 参考文献 】
  • [1]Osteryoung KW, Nunnari J: The division of endosymbiotic organelles. Science 2003, 302(5651):1698-1704.
  • [2]Karbowski M, Youle RJ: Dynamics of mitochondrial morphology in healthy cells and during apoptosis. Cell Death Differ 2003, 10(8):870-880.
  • [3]Scott I, Youle RJ: Mitochondrial fission and fusion. Essays Biochem 2010, 47:85-98.
  • [4]Johnson LV, Walsh ML, Chen LB: Localization of mitochondria in living cells with rhodamine 123. Proceedings of the National Academy of Science, USA 1980, 77:990-994.
  • [5]Nunnari J, Marshall W, Straight A, Murray A, Sedat JW, Walter P: Mitochondrial transmission during mating in S. cerevisiae is determined by mitochondrial fusion and fission and the intramitochondrial segregation of mtDNA. Molecular Biology of the Cell 1997, 8(7):1233-1242.
  • [6]Chen H, Chan DC: Mitochondrial dynamics–fusion, fission, movement, and mitophagy–in neurodegenerative diseases. Human Molecular Genetics 2009, 18(R2):R169-R176.
  • [7]Detmer SA, Chan DC: Functions and dysfunctions of mitochondrial dynamics. Nat Rev Mol Cell Biol 2007, 8(11):870-879.
  • [8]Karbowski M, Lee YJ, Gaume B, Jeong SY, Frank S, Nechushtan A, Santel A, Fuller M, Smith CL, Youle RJ: Spatial and temporal association of Bax with mitochondrial fission sites, Drp1, and Mfn2 during apoptosis. J Cell Biol 2002, 159(6):931-938.
  • [9]Frank S, Gaume B, Bergmann-Leitner ES, Leitner W, Robert EG, Catez F, Smith CL, Youle RJ: The role of dynamin-related protein 1, a mediator of mitochondrial fission, in apoptosis. Developmental Cell 2001, 1:515-525.
  • [10]Jin C, Reed JC: Yeast and apoptosis. Nat Rev Mol Cell Biol 2002, 3(6):453-459.
  • [11]Zuchner S, Mersiyanova IV, Muglia M, Bissar-Tadmouri N, Rochelle J, Dadali EL, Zappia M, Nelis E, Patitucci A, Senderek J, Parman Y, Evgrafov O, Jonghe PD, Takahashi Y, Tsuji S, Pericak-Vance MA, Quattrone A, Battologlu E, Polyakov AV, Timmerman V, Schroder JM, Vance JM: Mutations in the mitochondrial GTPase mitofusin 2 cause Charcot-Marie-Tooth neuropathy type 2A. Nat Genet 2004, 36(5):449-451. Epub 2004 Apr 2004
  • [12]Delettre C, Lenaers G, Griffoin J-M, Gigarel N, Lorenzo C, Belenguer P, Pelloquin L, Grosgeorge J, Turc-Carel C, Perret E, Astarie-Dequeker C, Lasquellec L, Arnaud B, Ducommun B, Kaplan J, Hamel CP: Nuclear gene OPA1, encoding a mitochondrial dynamin-related protein, is mutated in dominant optic atrophy. Nature Genetics 2000, 26(2):207-210.
  • [13]Alexander C, Votruba M, Pesch UEA, Thiselton DL, Mayer S, Moore A, Rodriguez M, Kellner U, Leo-Kottler B, Auburger G, Bhattacharya SS, Wissinger B: OPA1, encoding a dynamin-related GTPase, is mutated in autosomal dominant optic atrophy linked to chromosome 3q28. Nature Genetics 2000, 26(2):211-215.
  • [14]Otsuga D, Keegan BR, Brisch E, Thantcher JW, Hermann GJ, Bleazard W, Shaw J: The dynamin GTPase, Dnm1p, controls mitochondrial morphology in yeast. Journal of Cell Biology 1998, 143:333-349.
  • [15]Bleazard W, McCaffery JM, King EJ, Bale S, Mozdy A, Tieu Q, Nunnari J, Shaw JM: The dynamin-related GTPases, Dnm1, regulates mitochondrial fission in yeast. Nature Cell Biology 1999, 1:298-304.
  • [16]Sesaki H, Jensen RE: Division versus fusion: Dnm1p and Fzo1p antagonistically regulate mitochondrial shape. Journal of Cell Biology 1999, 147(4):699-706.
  • [17]Shaw JM, Nunnari J: Mitochondrial dynamics and division in budding yeast. Trends in Cell Biology 2002, 12(4):178-184.
  • [18]Osteryoung KW: Organelle fission in eukaryotes. Curr Opin Microbiol 2001, 4(6):639-646.
  • [19]Sever S: Dynamin and endocytosis. Curr Opin Cell Biol 2002, 14(4):463-467.
  • [20]Wienke DC, Knetsch MLW, Neuhaus EM, Reedy MC, Manstein DJ: Disruption of a Dynamin Homologue Affects Endocytosis, Organelle Morphology, and Cytokinesis in Dictyostelium discoideum. Mol Biol Cell 1999, 10(1):225-243.
  • [21]Rai A, Nothe H, Tzvetkov N, Korenbaum E, Manstein D: Dictyostelium dynamin B modulates cytoskeletal structures and membranous organelles. Cellular and Molecular Life Sciences 2011, 68:2751-2767.
  • [22]Scott SV, Cassidy-Stone A, Meeusen SL, Nunnari J: Staying in aerobic shape: how the structural integrity of mitochondria and mitochondrial DNA is maintained. Curr Opin Cell Biol 2003, 15(4):482-488.
  • [23]Gilson PR, Yu X-C, Hereld D, Barth C, Savage A, Kiefel BR, Lay S, Fisher PR, Margolin W, Beech PL: Two Dictyostelium Orthologs of the Prokaryotic Cell Division Protein FtsZ Localize to Mitochondria and Are Required for the Maintenance of Normal Mitochondrial Morphology. Eukaryotic Cell 2003, 2(6):1315-1326.
  • [24]Fields SD AQ, Heuser J, Clarke M: Mitochondrial membrane dynamics are altered in cluA- mutants of Dictyostelium. Journal of Muscle Research and Cell Motility 2002, 23:829-838.
  • [25]Torija P, Vicente JJ, Rodrigues TB, Robles A, Cerdan S, Sastre L, Calvo RM, Escalante R: Functional genomics in Dictyostelium: MidA, a new conserved protein, is required for mitochondrial function and development. Journal of Cell Science 2006, 119(6):1154-1164.
  • [26]Fields SD, Conrad MN, Clarke M: The S. cerevisiae CLU1 and D. discoideum cluA genes are functional homologues that influence mitochondrial morphology and distribution. Journal of Cell Science 1998, 111(12):1717-1727.
  • [27]Zhu Q, Hulen D, Liu T, Clarke M: The cluA- mutant of Dictyostelium identifies a novel class of proteins required for dispersion of mitochondria. Proceedings of the National Academy of Sciences 1997, 94(14):7308-7313.
  • [28]Dictybasewww.dictybase.org webcite
  • [29]Fey P, Kowal AS, Gaudet P, Pilcher KE, Chisholm RL: Protocols for growth and development of Dictyostelium discoideum. 2007, 2(6):1307-1316.
  • [30]Poot M, Zhang YZ, Krämer JA, Wells KS, Jones LJ, Hanzel DK, Lugade AG, Singer VL, Haugland RP: Analysis of mitochondrial morphology and function with novel fixable fluorescent stains. Journal of Histochemistry & Cytochemistry 1996, 44(12):1363-1372.
  • [31]Carilla-Latorre S, Gallardo ME, Annesley SJ, Calvo-Garrido J, Grana O, Accari SL, Smith PK, Valencia A, Garesse R, Fisher PR, Escalante R: MidA is a putative methyltransferase that is required for mitochondrial complex I function. Journal of Cell Science 2010, 123(10):1674-1683.
  • [32]UniProt CT: Reorganizing the protein space at the Universal Protein Resource (UniProt). Nucleic Acids Res 2012, 40:D71-D75.
  • [33]Logan DC, Scott I, Tobin AK: The genetic control of plant mitochondrial morphology and dynamics. The Plant Journal 2003, 36(4):500-509.
  • [34]Cox RT, Spradling AC: clueless, a conserved Drosophila gene required fro mitochondrial subcellular localization, interacts genetically with parkin. Disease Models and Mechanisms 2009, 2(9–10):490-499.
  • [35]Motley AM, Hettema EH: Yeast peroxisomes multiply by growth and division. The Journal of Cell Biology 2007, 178(3):399-410.
  • [36]Dohm JA, Lee SJ, Hardwick JM, Hill RB, Gittis AG: Cytosolic domain of the human mitochondrial fission protein fis1 adopts a TPR fold. Proteins: Structure, Function, and Bioinformatics 2004, 54(1):153-156.
  • [37]Motley AM, Ward GP, Hettema EH: Dnm1p-dependent peroxisome fission requires Caf4p, Mdv1p and Fis1p. Journal of Cell Science 2008, 121(10):1633-1640.
  文献评价指标  
  下载次数:69次 浏览次数:52次