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Article

Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids

  • Authors:
    • Tomáš Olejár
    • David Pajuelo‑Reguera
    • Lukáš Alán
    • Andrea Dlasková
    • Petr Ježek
  • View Affiliations / Copyright

    Affiliations: Department of Membrane Transport Biophysics, Institute of Physiology, The Czech Academy of Sciences, Prague 14220, Czech Republic
  • Pages: 5185-5190
    |
    Published online on: July 16, 2015
       https://doi.org/10.3892/mmr.2015.4085
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Abstract

Oligomer aggregation of green-to-red photoconvertible fluorescent protein Eos (EosFP) is a natural feature of the wild‑type variant. The aim of the present study was to follow up mitochondrial nucleoid behavior under natural conditions of living cells transfected with mitochondrial single‑strand DNA‑binding protein (mtSSB) conjugated with EosFP. HEPG2 and SH‑SY5Y cells were subjected to lentiviral transfection and subsequently immunostained with anti‑DNA, anti‑transcription factor A, mitochondrial (TFAM) or anti‑translocase of the inner membrane 23 antibodies. Fluorescent microscopy, conventional confocal microscopy, superresolution biplane fluorescence photo-activation localization microscopy and direct stochastic optical reconstruction microscopy were used for imaging. In the two cell types, apparent couples of equally‑sized mtSSB‑EosFP‑visualized dots were observed. During the time course of the ongoing transfection procedure, however, a small limited number of large aggregates of mtSSB‑EosFP‑tagged protein started to form in the cells, which exhibited a great co‑localization with the noted coupled positions. Antibody staining and 3D immunocytochemistry confirmed that nucleoid components such as TFAM and DNA were co‑localized with these aggregates. Furthermore, the observed reduction of the mtDNA copy number in mtSSB‑EosFP‑transfected cells suggested a possible impairment of nucleoid function. In conclusion, the present study demonstrated that coupled nucleoids are synchronized by mtSSB‑EosFP overexpression and visualized through their equal binding capacity to mtSSB‑EosFP‑tagged protein. This observation suggested parallel replication and transcription activity of nucleoid couples native from a parental one. Preserved coupling in late stages of artificial EosFP‑mediated aggregation of tagged proteins suggested a rational manner of mitochondrial branching that may be cell-type specifically dependent on hierarchical nucleoid replication.
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1 

Brown TA, Tkachuk AN, Shtengel G, Kopek BG, Bogenhagen DF, Hess HF and Clayton DA: Superresolution fluorescence imaging of mitochondrial nucleoids reveals thein spatial range, limits and membrane interaction. Mol Cell Biol. 31:4994–5010. 2011. View Article : Google Scholar : PubMed/NCBI

2 

Bogenhagen DF: Mitochondrial DNA nucleoid structure. Biochim Biophys Acta. 1819:914–920. 2012. View Article : Google Scholar

3 

Ngo HB, Kaiser JT and Chan DC: The mitochondrial transcription and packaging factor Tfam imposes a U-turn on mitochondrial DNA. Nat Struct Mol Biol. 18:1290–1296. 2011. View Article : Google Scholar : PubMed/NCBI

4 

Scarpulla RC: Transcriptional paradigms in mammalian mitochondrial biogenesis and function. Physiol Rev. 88:611–638. 2008. View Article : Google Scholar : PubMed/NCBI

5 

Falkenberg M, Larsson NG and Gustafsson CM: DNA replication and transcription in mammalian mitochondria. Annu Rev Biochem. 76:679–699. 2007. View Article : Google Scholar : PubMed/NCBI

6 

Campbell CT, Kolesar JE and Kaufman BA: Mitochondrial transcription factor A regulates mitochondrial transcription initiation, DNA packaging and genome copy number. Biochim Biophys Acta. 1819:921–929. 2012. View Article : Google Scholar : PubMed/NCBI

7 

Gauthier BR, Wiederkehr A, Baquié M, Dai C, Powers AC, Kerr-Conte J, Pattou F, MacDonald RJ, Ferrer J and Wollheim CB: PDX1 deficiency causes mitochondrial dysfunction and defective insulin secretion through TFAM suppression. Cell Metab. 10:110–118. 2009. View Article : Google Scholar : PubMed/NCBI

8 

Shi CM, Xu GF, Yang L, Fu ZY, Chen L, Fu HL, Shen YH, Zhu L, Ji CB and Guo XR: Overexpression of TFAM protects 3T3-L1 adipocytes from NYGGF4 (PID1) overexpression-induced insulin resistance and mitochondrial dysfunction. Cell Biochem Biophys. 66:489–497. 2013. View Article : Google Scholar : PubMed/NCBI

9 

Kaguni LS: DNA polymerase gamma, the mitochondrial replicase. Annu Rev Biochem. 73:293–320. 2004. View Article : Google Scholar : PubMed/NCBI

10 

Ruhanen H, Borrie S, Szabadkai G, Tyynismaa H, Jones AW, Kang D, Taanman JW and Yasukawa T: Mitochondrial single-stranded DNA binding protein is required for maintenance of mitochondrial DNA and 7S DNA but is not required for mitochondrial nucleoid organisation. Biochim Biophys Acta. 1803:931–939. 2010. View Article : Google Scholar : PubMed/NCBI

11 

Van Tuyle GC and Pavco PA: The rat liver mitochondrial DNA-protein complex: Displaced single strands of replicative intermediates are protein coated. J Cell Biol. 100:251–257. 1985. View Article : Google Scholar : PubMed/NCBI

12 

Hoke GD, Pavco PA, Ledwith BJ and Van Tuyle GC: Structural and functional studies of the rat mitochondrial single strand DNA binding protein P16. Arch Biochem Biophys. 282:116–124. 1990. View Article : Google Scholar : PubMed/NCBI

13 

Korhonen JA, Gaspari M and Falkenberg M: TWINKLE Has 5′->3′ DNA helicase activity and is specifically stimulated by mitochondrial single-stranded DNA-binding protein. J Biol Chem. 278:48627–48632. 2003. View Article : Google Scholar : PubMed/NCBI

14 

Milenkovic D, Matic S, Kühl I, Ruzzenente B, Freyer C, Jemt E, Park CB, Falkenberg M and Larsson NG: TWINKLE is an essential mitochondrial helicase required for synthesis of nascent D-loop strands and complete mtDNA replication. Hum Mol Genet. 22:1983–1993. 2013. View Article : Google Scholar : PubMed/NCBI

15 

Blomain ES and McMahon SB: Dynamic regulation of mitochondrial transcription as a mechanism of cellular adaptation. Biochim Biophys Acta. 1819:1075–1079. 2012. View Article : Google Scholar : PubMed/NCBI

16 

Asin-Cayuela J and Gustafsson CM: Mitochondrial transcription and its regulation in mammalian cells. Trends Biochem Sci. 32:111–117. 2007. View Article : Google Scholar : PubMed/NCBI

17 

Tauber J, Dlasková A, Santorová J, Smolková K, Alán L, Spaček T, Plecitá-Hlavatá L, Jabůrek M and Ježek P: Distribution of mitochondrial nucleoids upon mitochondrial network fragmentation and network reintegration in HEPG2 cells. Int J Biochem Cell Biol. 45:593–603. 2013. View Article : Google Scholar

18 

Li Z, Shen J, Chen Y, Pan J, Zeng H, Fang H, Ye Z, Zeng C, Zhang R and Cai D: Mitochondrial genome sequencing of chondrocytes in osteoarthritis by human mitochondria RT2 Profiler™ PCR array. Mol Med Rep. 6:39–44. 2012.PubMed/NCBI

19 

Piao L, Han Y and Li D: Correlation study on adiponectin gene SNP45 and long-term oxidative stress in patients with diabetes and carotid atherosclerosis. Exp Ther Med. 8:707–712. 2014.PubMed/NCBI

20 

Grzybowska-Szatkowska L and Slaska B: Mitochondrial DNA and carcinogenesis (review). Mol Med Rep. 6:923–930. 2012.PubMed/NCBI

21 

Li WH and Zheng G: Photoactivatable fluorophores and techniques for biological imaging applications. Photochem Photobiol Sci. 11:460–471. 2012. View Article : Google Scholar : PubMed/NCBI

22 

Hedde PN and Nienhaus GU: Super-resolution localization microscopy with photoactivatable fluorescent marker proteins. Protoplasma. 251:349–362. 2014. View Article : Google Scholar

23 

Nienhaus GU, Nienhaus K, Hölzle A, Ivanchenko S, Renzi F, Oswald F, Wolff M, Schmitt F, Röcker C, Vallone B, et al: Photoconvertible fluorescent protein EosFP: Biophysical properties and cell biology applications. Photochem Photobiol. 82:351–358. 2006. View Article : Google Scholar : PubMed/NCBI

24 

Mlodzianoski MJ, Schreiner JM, Callahan SP, Smolková K, Dlasková A, Santorová J, Ježek P and Bewersdorf J: Sample drift correction in 3D fluorescence photoactivation localization microscopy. Opt Express. 19:15009–15019. 2011. View Article : Google Scholar : PubMed/NCBI

25 

Alán L, Špaček T, Zelenka J, Tauber J, Berková Z, Zacharovová K, Saudek F and Ježek P: Assessment of mitochondrial DNA as an indicator of islet quality: An example in Goto Kakizaki rats. Transplant Proc. 43:3281–3284. 2011. View Article : Google Scholar : PubMed/NCBI

26 

Korr H, Thorsten Rohde H, Benders J, Dafotakis M, Grolms N and Schmitz C: Neuron loss during early adulthood following prenatal low-dose X-irradiation in the mouse brain. Int J Radiat Biol. 77:567–580. 2001. View Article : Google Scholar : PubMed/NCBI

27 

Arakaki N, Nishihama T, Kohda A, Owaki H, Kuramoto Y, Abe R, Kita T, Suenaga M, Himeda T, Kuwajima M, et al: Regulation of mitochondrial morphology and cell survival by Mitogenin I and mitochondrial single-stranded DNA binding protein. Biochim Biophys Acta. 1760:1364–1372. 2006. View Article : Google Scholar : PubMed/NCBI

28 

Iborra FJ, Kimura H and Cook PR: The functional organization of mitochondrial genomes in human cells. BMC Biol. 2:92004. View Article : Google Scholar : PubMed/NCBI

29 

Ban-Ishihara R, Ishihara T, Sasaki N, Mihara K and Ishihara N: Dynamics of nucleoid structure regulated by mitochondrial fission contributes to cristae reformation and release of cytochrome c. Proc Natl Acad Sci USA. 110:11863–11868. 2013. View Article : Google Scholar : PubMed/NCBI

30 

Yang C, Curth U, Urbanke C and Kang C: Crystal structure of human mitochondrial single-stranded DNA binding protein at 2.4 A resolution. Nat Struct Biol. 4:153–157. 1997. View Article : Google Scholar : PubMed/NCBI

31 

Antony E, Weiland E, Yuan Q, Manhart CM, Nguyen B, Kozlov AG, McHenry CS and Lohman TM: Multiple C-terminal tails within a single E. coli SSB homotetramer coordinate DNA replication and repair. J Mol Biol. 425:4802–4819. 2013. View Article : Google Scholar : PubMed/NCBI

32 

Rajala N, Gerhold JM, Martinsson P, Klymov A and Spelbrink JN: Replication factors transiently associate with mtDNA at the mitochondrial inner membrane to facilitate replication. Nucleic Acids Res. 42:952–967. 2014. View Article : Google Scholar :

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Copy and paste a formatted citation
Spandidos Publications style
Olejár T, Pajuelo‑Reguera D, Alán L, Dlasková A and Ježek P: Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids. Mol Med Rep 12: 5185-5190, 2015.
APA
Olejár, T., Pajuelo‑Reguera, D., Alán, L., Dlasková, A., & Ježek, P. (2015). Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids. Molecular Medicine Reports, 12, 5185-5190. https://doi.org/10.3892/mmr.2015.4085
MLA
Olejár, T., Pajuelo‑Reguera, D., Alán, L., Dlasková, A., Ježek, P."Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids". Molecular Medicine Reports 12.4 (2015): 5185-5190.
Chicago
Olejár, T., Pajuelo‑Reguera, D., Alán, L., Dlasková, A., Ježek, P."Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids". Molecular Medicine Reports 12, no. 4 (2015): 5185-5190. https://doi.org/10.3892/mmr.2015.4085
Copy and paste a formatted citation
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Spandidos Publications style
Olejár T, Pajuelo‑Reguera D, Alán L, Dlasková A and Ježek P: Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids. Mol Med Rep 12: 5185-5190, 2015.
APA
Olejár, T., Pajuelo‑Reguera, D., Alán, L., Dlasková, A., & Ježek, P. (2015). Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids. Molecular Medicine Reports, 12, 5185-5190. https://doi.org/10.3892/mmr.2015.4085
MLA
Olejár, T., Pajuelo‑Reguera, D., Alán, L., Dlasková, A., Ježek, P."Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids". Molecular Medicine Reports 12.4 (2015): 5185-5190.
Chicago
Olejár, T., Pajuelo‑Reguera, D., Alán, L., Dlasková, A., Ježek, P."Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids". Molecular Medicine Reports 12, no. 4 (2015): 5185-5190. https://doi.org/10.3892/mmr.2015.4085
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