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Silencing of NbBTF3 results in developmental defects and disturbed gene expression in chloroplasts and mitochondria of higher plants

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Abstract

BTF3 (βNAC) was originally isolated as a general transcription factor required for RNA polymerase II-dependent transcription, and later found to be a β-subunit of nascent-polypeptide-associated complex that has been implicated in regulating protein localization during translation. In this study, virus-induced gene silencing of NbBTF3 encoding a Nicotiana benthamiana homolog of human BTF3 caused leaf yellowing and abnormal leaf morphology without altering the overall growth of the plant. The NbBTF3 gene is constitutively expressed and the NbBTF3–GFP fusion protein is primarily targeted to the nucleus. At the cellular level, downregulation of NbBTF3 expression reduced the chloroplast sizes and chlorophyll contents. The affected cells produced excessive amounts of reactive oxygen species. Furthermore, the transcript level of various plastid- and mitochondria-encoded genes was severely reduced in the NbBTF3-depleted leaf cells. These findings indicate that depletion of NbBTF3 activity preferentially affected development and/or physiology of chloroplasts and mitochondria in plants, possibly by hampering efficient translocation of the nascent organellar proteins into the organelles.

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Abbreviations

BTF3:

β-Subunit of NAC

NAC:

Nascent polypeptide-associated complex

NEP:

Nucleus-encoded phage-type RNA polymerases

NLS:

Nuclear localization signal

PEP:

Plastid-encoded multimeric RNA polymerase

ROS:

Reactive oxygen species

TMRM:

Tetramethylrhodamine methyl ester

TRV:

Tobacco rattle virus

VIGS:

Virus-induced gene silencing

References

  • Allison LA (2000) The role of sigma factors in plastid transcription. Biochimie 82:537–548

    Article  PubMed  CAS  Google Scholar 

  • Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

    Article  PubMed  CAS  Google Scholar 

  • Baba K, Schmidt J, Espinosa-Ruiz A, Villarejo A, Shiina T, Gardestrom P, Sane AP, Bhalerao RP (2004) Organellar gene transcription and early seedling development are affected in the rpoT; 2 mutant of Arabidopsis. Plant J 38:38–48

    Article  PubMed  CAS  Google Scholar 

  • Beatrix B, Sakai H, Wiedmann M (2000) The alpha and beta subunit of the nascent polypeptide-associated complex have distinct functions. J Biol Chem 275:37838–37845

    Article  PubMed  CAS  Google Scholar 

  • Bethke PC, Jones RL (2001) Cell death of barley aleurone protoplasts is mediated by reactive oxygen species. Plant J 25:19–29

    Article  PubMed  CAS  Google Scholar 

  • Bloss TA, Wilze ES, Rothman JH (2003) Suppression of CED-3-independent apoptosis by mitochondrial βNAC in Caenorhabditis elegans. Nature 424:1066–1071

    Article  PubMed  CAS  Google Scholar 

  • Braat AK, Yan N, Arn E, Harrison D, Macdonald PM (2004) Localization-dependent oskar protein accumulation: control after the initiation of translation. Dev Cell 7:125–131

    Article  PubMed  CAS  Google Scholar 

  • Brockstedt E, Otto A, Rickers A, Bommert K, Wittmann-Liebold B (1999) Preparative high-resolution two-dimensional electrophoresis enables the identification of RNA polymerase B transcription factor B as an apoptosis-associated protein in the human BL60-2 Burkitt lymphoma cell line. J Protein Chem 18:225–231

    Article  PubMed  CAS  Google Scholar 

  • Cho HS, Lee SS, Kim KD, Kim SJ, Hwang I, Lim JS, Park YI, Pai HS (2004) DNA gyrase is involved in chloroplast nucleoid partitioning. Plant Cell 16:2665–2682

    Article  PubMed  CAS  Google Scholar 

  • Deng JM, Behringer RR (1995) An insertional mutation in the BTF3 transcription factor gene leads to an early postimplantation lethality in mice. Transgenic Res 4:264–269

    Article  PubMed  CAS  Google Scholar 

  • Franke J, Reimann B, Hartman E, Köhler M, Wiedmann B (2001) Evidence for a nuclear passage of nascent polypeptide-associated complex subunits in yeast. J Cell Sci 114:2641–2648

    PubMed  CAS  Google Scholar 

  • Freire MA (2005) Translation initiation factor (iso) 4E interacts with BTF3, the β subunit of the nascent polypeptide-associated complex. Gene 345:271–277

    Article  PubMed  CAS  Google Scholar 

  • Fujiwara M, Nagashima A, Kanamaru K, Tanaka K, Takahashi H (2000) Three new nuclear genes, sigD, sigE and sigF, encoding putative plastid RNA polymerase sigma factors in Arabidopsis thaliana. FEBS Lett 481:47–52

    Article  PubMed  CAS  Google Scholar 

  • Fünfschilling U, Rospert S (1999) Nascent polypeptide-associated complex stimulates protein import into yeast mitochondria. Mol Biol Cell 10:3289–3299

    PubMed  Google Scholar 

  • George R, Beddoe T, Landl K, Lithgow T (1998) The yeast nascent polypeptide-associated complex initiates protein targeting to mitochondria in vivo. Proc Natl Acad Sci USA 95:2296–2301

    Article  PubMed  CAS  Google Scholar 

  • George R, Walsh P, Beddoe T, Lithgow T (2002) The nascent polypeptide-associated complex (NAC) promotes interaction of ribosomes with the mitochondrial surface in vivo. FEBS Lett 516:213–216

    Article  PubMed  CAS  Google Scholar 

  • Green DR, Reed JC (1998) Mitochondria and apoptosis. Science 281:1309–1312

    Article  PubMed  CAS  Google Scholar 

  • Greenleaf AL, Kelly JL, Lehman IR (1986) Yeast RPO41 gene product is required for transcription and maintenance of the mitochondrial genome. Proc Natl Acad Sci USA 83:3391–3394

    Article  PubMed  CAS  Google Scholar 

  • Hajdukiewicz PT, Allison LA, Maliga P (1997) The two RNA polymerases encoded by the nuclear and the plastid compartments transcribe distinct groups of genes in tobacco plastids. EMBO J 16:4041–4048

    Article  PubMed  CAS  Google Scholar 

  • Hedtke B, Borner T, Weihe A (1997) Mitochondrial and chloroplast phage-type RNA polymerases in Arabidopsis. Science 277:809–811

    Article  PubMed  CAS  Google Scholar 

  • Hedtke B, Borner T, Weihe A (2000) One RNA polymerase serving two genomes. EMBO Rep 1:435–440

    Article  PubMed  CAS  Google Scholar 

  • Hu GZ, Ronne H (1994) Yeast BTF3 protein is encoded by duplicated genes and inhibits the expression of some genes in vivo. Nucleic Acids Res 22:2740–2743

    Article  PubMed  CAS  Google Scholar 

  • Ishizaki Y, Tsunoyama Y, Hatano K, Ando K, Kato K, Shinmyo A, Kobori M, Takeba G, Nakahira Y, Shiina T (2005) A nuclear-encoded sigma factor, Arabidopsis SIG6, recognizes sigma-70 type chloroplast promoters and regulates early chloroplast development in cotyledons. Plant J 42:133–144

    Article  PubMed  CAS  Google Scholar 

  • Kim YS, Lee JY, Cho HS, Lee SS, Ha HJ, Kim S, Choi D, Pai HS (2005) Inactivation of an organellar glutamyl- and seryl-tRNA synthetase leads to a developmental arrest of chloroplasts and mitochondria in higher plants. J Biol Chem 280:37098–37106

    Article  PubMed  CAS  Google Scholar 

  • Lam E, Kato N, Lawton M (2001) Programmed cell death, mitochondria and the plant hypersensitive response. Nature 411:848–853

    Article  PubMed  CAS  Google Scholar 

  • Lauring B, Sakai H, Kreibich G, Wiedmann M (1995) Nascent polypeptide-associated complex protein prevents mistargeting of nascent chains to the endoplasmic reticulum. Proc Natl Acad Sci USA 92:5411–5415

    Article  PubMed  CAS  Google Scholar 

  • Lee YJ, Kim DH, Kim YW, Hwang I (2001) Identification of a signal that distinguishes between the chloroplast outer envelope membrane and the endomembrane system in vivo. Plant Cell 13:2175–2190

    Article  PubMed  CAS  Google Scholar 

  • Magee AM, Kavanagh TA (2002) Plastid genes transcribed by the nucleus-encoded plastid RNA polymerase show increased transcript accumulation in transgenic plants expressing a chloroplast-localized phage T7 RNA polymerase. J Exp Bot 53:2341–2349

    Article  PubMed  CAS  Google Scholar 

  • Maliga P (1998) Two plastid RNA polymerases in higher plants: an evolving story. Trends Plant Sci 3:4–6

    Article  Google Scholar 

  • Markesich DC, Gajewski KM, Nazimiec ME, Beckingham K (2000) bicaudal encodes the Drosophila beta NAC homolog a component of the ribosomal translational machinery. Development 127:559–572

    PubMed  CAS  Google Scholar 

  • Maxwell DP, Wang Y, McIntosh L (1999) The alternative oxidase lowers mitochondrial reactive oxygen production in plant cells. Proc Natl Acad Sci USA 96:8271–8276

    Article  PubMed  CAS  Google Scholar 

  • Moreau A, Yotov WV, Glorieux FH, St-Arnaud R (1998) Bone-specific expression of the alpha chain of the nascent polypeptide-associated complex a coactivator potentiating c-Jun-mediated transcription. Mol Cell Biol 18:1312–1321

    PubMed  CAS  Google Scholar 

  • Parthun MR, Mangus DA, Jaehning JA (1992) The EGD1 product, a yeast homolog of human BTF3, may be involved in GAL4 DNA binding. Mol Cell Biol 12:5683–5689

    PubMed  CAS  Google Scholar 

  • Quélo I, Gauthier C, Hannigan GE, Dedhar S, St-Arnaud R (2004) Integrin-linked kinase regulates the nuclear entry of the c-Jun coactivator alpha-NAC and its coactivation potency. J Biol Chem 279:43893–43899

    Article  PubMed  Google Scholar 

  • Ratcliff F, Martin-Hernansez AM, Baulcombe DC (2001) Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J 25:237–245

    Article  PubMed  CAS  Google Scholar 

  • Rospert S, Dubaquié Y, Gautschi M (2002) Nascent-polypeptide-associated complex. Cell Mol Life Sci 59:1632–1639

    Article  PubMed  CAS  Google Scholar 

  • Silhavy D, Maliga P (1998) Mapping of promoters for the nucleus-encoded plastid RNA polymerase (NEP) in the iojap maize mutant. Curr Genet 33:340–344

    Article  PubMed  CAS  Google Scholar 

  • Wang S, Sakai H, Kreibich G, Wiedmann M (1995) NAC covers ribosome-associated nascent chains thereby forming a protective environment for regions of nascent chains just emerging from the peptidyl transferase center. J Cell Biol 130:519–528

    Article  PubMed  CAS  Google Scholar 

  • Wiedmann B, Sakai H, Davis TA, Wiedmann M (1994) A protein complex required for signal-sequence-specific sorting and translocation. Nature 370:434–440

    Article  PubMed  CAS  Google Scholar 

  • Zhang H, Huang HM, Carson RC, Mahmood J, Thomas HM, Gibson GE (2001) Assessment of membrane potentials of mitochondrial populations in living cells. Anal Biochem 298:170–180

    Article  PubMed  CAS  Google Scholar 

  • Zheng XM, Moncollin V, Egly JM, Chambon P (1987) A general transcription factor forms a stable complex with RNA polymerase B (II). Cell 50:361–368

    Article  PubMed  CAS  Google Scholar 

  • Zheng XM, Black D, Chambon P, Egly JM (1990) Sequencing and expression of complementary DNA for the general transcription factor BTF3. Nature 344:556–559

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This research was supported by the grants from the Molecular and Cellular BioDiscovery Program and KOSEF Basic Research Program (MOST, Korea), and from the BioGreen21 Program (RDA, Korea).

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Author notes

  1. Kyoung-Sil Yang

    Present address: Environmental Biotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, Taejon, 305-333, South Korea

Authors and Affiliations

  1. Laboratory of Plant Genomics, Korea Research Institute of Bioscience and Biotechnology, Taejon, 305-333, South Korea

    Kyoung-Sil Yang, Un-Ho Jin & Sang Sook Lee

  2. Department of Biology, Yonsei University, Seoul, 120-749, South Korea

    Hee-Seung Kim, Jong-A Park & Hyun-Sook Pai

  3. Department of Horticulture, Chungnam University, Taejon, 305-764, South Korea

    Kyoung-Sil Yang & Yong Pyo Lim

  4. Department of Genetics, University of Wisconsin, Madison, WI, 53706-1574, USA

    Sang Sook Lee

  5. Department of Biological Science, Sungkyunkwan University, Suwon, Kyonggi-do, 440-746, South Korea

    Un-Ho Jin

  6. Section of Plant Biology, Department of Plant Sciences, University of California, Davis, CA, 95616, USA

    Jong-A Park

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Correspondence to Hyun-Sook Pai.

Additional information

K.-S. Yang and H.-S. Kim contributed equally to this work.

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425_2006_453_MOESM1_ESM.ppt

Expression of the NbBTF3 gene. Semiquantitative RT-PCR analysis was carried out with RNA isolated from N. benthamiana plants using the NbBTF3– specific primers. As a control, the transcript level of actin was examined. a Expression in various tissues of N. benthamiana plants. b Expression in response to various hormones. c Expression in response to the heat (42°C) treatment for the indicated times. d Expression in response to the NaCl treatment (0–400 mM) for 24 h. e Light-stimulated expression. Seedlings were grown under normal light conditions (16 h light/8 h dark) (light), under dark conditions (dark), or under dark conditions followed by a transfer to light for 1 h (dark→light) (PPT 84 kb)

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Yang, KS., Kim, HS., Jin, UH. et al. Silencing of NbBTF3 results in developmental defects and disturbed gene expression in chloroplasts and mitochondria of higher plants. Planta 225, 1459–1469 (2007). https://doi.org/10.1007/s00425-006-0453-3

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  • DOI: https://doi.org/10.1007/s00425-006-0453-3

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