The Bicoid Stability Factor Controls Polyadenylation and Expression of Specific Mitochondrial mRNAs in
The bicoid stability factor (BSF) of Drosophila melanogaster has been reported to be present in the cytoplasm, where it stabilizes the maternally contributed bicoid mRNA and binds mRNAs expressed from early zygotic genes. BSF may also have other roles, as it is ubiquitously expressed and essential for survival of adult flies. We have performed immunofluorescence and cell fractionation analyses and show here that BSF is mainly a mitochondrial protein. We studied two independent RNAi knockdown fly lines and report that reduced BSF protein levels lead to a severe respiratory deficiency and delayed development at the late larvae stage. Ubiquitous knockdown of BSF results in a severe reduction of the polyadenylation tail lengths of specific mitochondrial mRNAs, accompanied by an enrichment of unprocessed polycistronic RNA intermediates. Furthermore, we observed a significant reduction in mRNA steady state levels, despite increased de novo transcription. Surprisingly, mitochondrial de novo translation is increased and abnormal mitochondrial translation products are present in knockdown flies, suggesting that BSF also has a role in coordinating the mitochondrial translation in addition to its role in mRNA maturation and stability. We thus report a novel function of BSF in flies and demonstrate that it has an important intra-mitochondrial role, which is essential for maintaining mtDNA gene expression and oxidative phosphorylation.
Vyšlo v časopise:
The Bicoid Stability Factor Controls Polyadenylation and Expression of Specific Mitochondrial mRNAs in. PLoS Genet 7(10): e32767. doi:10.1371/journal.pgen.1002324
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1002324
Souhrn
The bicoid stability factor (BSF) of Drosophila melanogaster has been reported to be present in the cytoplasm, where it stabilizes the maternally contributed bicoid mRNA and binds mRNAs expressed from early zygotic genes. BSF may also have other roles, as it is ubiquitously expressed and essential for survival of adult flies. We have performed immunofluorescence and cell fractionation analyses and show here that BSF is mainly a mitochondrial protein. We studied two independent RNAi knockdown fly lines and report that reduced BSF protein levels lead to a severe respiratory deficiency and delayed development at the late larvae stage. Ubiquitous knockdown of BSF results in a severe reduction of the polyadenylation tail lengths of specific mitochondrial mRNAs, accompanied by an enrichment of unprocessed polycistronic RNA intermediates. Furthermore, we observed a significant reduction in mRNA steady state levels, despite increased de novo transcription. Surprisingly, mitochondrial de novo translation is increased and abnormal mitochondrial translation products are present in knockdown flies, suggesting that BSF also has a role in coordinating the mitochondrial translation in addition to its role in mRNA maturation and stability. We thus report a novel function of BSF in flies and demonstrate that it has an important intra-mitochondrial role, which is essential for maintaining mtDNA gene expression and oxidative phosphorylation.
Zdroje
1. ManceboRZhouXShillinglawWHenzelWMacdonaldPM 2001 BSF binds specifically to the bicoid mRNA 3′ untranslated region and contributes to stabilization of bicoid mRNA. Mol Cell Biol 21 3462 3471 doi:10.1128/MCB.21.10.3462-3471.2001
2. De RenzisSElementoOTavazoieSWieschausEF 2007 Unmasking activation of the zygotic genome using chromosomal deletions in the Drosophila embryo. PLoS Biol 5 e117 doi:10.1371/journal.pbio.0050117
3. ChintapalliVRWangJDowJAT 2007 Using FlyAtlas to identify better Drosophila melanogaster models of human disease. Nat Genet 39 715 720 doi:10.1038/ng2049
4. SterkyFHRuzzenenteBGustafssonCMSamuelssonTLarssonN-G 2010 LRPPRC is a mitochondrial matrix protein that is conserved in metazoans. Biochemical and Biophysical Research Communications 398 759 764 doi:10.1016/j.bbrc.2010.07.019
5. LurinCAndrésCAubourgSBellaouiMBittonF 2004 Genome-wide analysis of Arabidopsis pentatricopeptide repeat proteins reveals their essential role in organelle biogenesis. Plant Cell 16 2089 2103 doi:10.1105/tpc.104.022236
6. MiliSShuHJZhaoYPiñol-RomaS 2001 Distinct RNP complexes of shuttling hnRNP proteins with pre-mRNA and mRNA: candidate intermediates in formation and export of mRNA. Mol Cell Biol 21 7307 7319 doi:10.1128/MCB.21.21.7307-7319.2001
7. TsuchiyaNFukudaHNakashimaKNagaoMSugimuraT 2004 LRP130, a single-stranded DNA/RNA-binding protein, localizes at the outer nuclear and endoplasmic reticulum membrane, and interacts with mRNA in vivo. Biochemical and Biophysical Research Communications 317 736 743 doi:10.1016/j.bbrc.2004.03.103
8. TopisirovicISiddiquiNLapointeVLTrostMThibaultP 2009 Molecular dissection of the eukaryotic initiation factor 4E (eIF4E) export-competent RNP. EMBO J 28 1087 1098 doi:10.1038/emboj.2009.53
9. LabialleSDayanGGayetLRigalDGambrelleJ 2004 New invMED1 element cis-activates human multidrug-related MDR1 and MVP genes, involving the LRP130 protein. Nucleic Acids Res 32 3864 3876 doi:10.1093/nar/gkh722
10. CooperMPQuLRohasLMLinJYangW 2006 Defects in energy homeostasis in Leigh syndrome French Canadian variant through PGC-1alpha/LRP130 complex. Genes Dev 20 2996 3009 doi:10.1101/gad.1483906
11. XuFMorinCMitchellGAckerleyCRobinsonBH 2004 The role of the LRPPRC (leucine-rich pentatricopeptide repeat cassette) gene in cytochrome oxidase assembly: mutation causes lowered levels of COX (cytochrome c oxidase) I and COX III mRNA. Biochem J 382 331 336 doi:10.1042/BJ20040469
12. SasarmanFBrunel-GuittonCAntonickaHWaiTShoubridgeEA 2010 LRPPRC and SLIRP interact in a ribonucleoprotein complex that regulates posttranscriptional gene expression in mitochondria. Mol Biol Cell 21 1315 1323 doi:10.1091/mbc.E10-01-0047
13. BooreJL 1999 Animal mitochondrial genomes. Nucleic Acids Res 27 1767 1780
14. MontoyaJOjalaDAttardiG 1981 Distinctive features of the 5′-terminal sequences of the human mitochondrial mRNAs. Nature 290 465 470
15. OjalaDMontoyaJAttardiG 1981 tRNA punctuation model of RNA processing in human mitochondria. Nature 290 470 474
16. TemperleyRJWydroMLightowlersRNChrzanowska-LightowlersZM 2010 Human mitochondrial mRNAs--like members of all families, similar but different. Biochim. Biophys. Acta 1797 1081 1085 doi:10.1016/j.bbabio.2010.02.036
17. TomeckiRDmochowskaAGewartowskiKDziembowskiAStepienPP 2004 Identification of a novel human nuclear-encoded mitochondrial poly(A) polymerase. Nucleic Acids Res 32 6001 6014 doi:10.1093/nar/gkh923
18. GohilVMNilssonRBelcher-TimmeCALuoBRootDE 2010 Mitochondrial and nuclear genomic responses to loss of LRPPRC expression. Journal of Biological Chemistry 285 13742 13747 doi:10.1074/jbc.M109.098400
19. FalkenbergMLarssonN-GGustafssonCM 2007 DNA replication and transcription in mammalian mitochondria. Annu Rev Biochem 76 679 699 doi:10.1146/annurev.biochem.76.060305.152028
20. GagliardiDStepienPPTemperleyRJLightowlersRNChrzanowska-LightowlersZMA 2004 Messenger RNA stability in mitochondria: different means to an end. Trends Genet 20 260 267 doi:10.1016/j.tig.2004.04.006
21. CoucheronDHNymarkMBreinesRKarlsenBOAndreassenM 2011 Characterization of mitochondrial mRNAs in codfish reveals unique features compared to mammals. Curr Genet 57 213 222 doi:10.1007/s00294-011-0338-2
22. BobrowiczAJLightowlersRNChrzanowska-LightowlersZ 2008 Polyadenylation and degradation of mRNA in mammalian mitochondria: a missing link? Biochem. Soc Trans 36 517 519 doi:10.1042/BST0360517
23. ColganDFManleyJL 1997 Mechanism and regulation of mRNA polyadenylation. Genes Dev 11 2755 2766
24. AphasizhevaIMaslovDWangXHuangLAphasizhevR 2011 Pentatricopeptide repeat proteins stimulate mRNA adenylation/uridylation to activate mitochondrial translation in trypanosomes. Mol Cell 42 106 117 doi:10.1016/j.molcel.2011.02.021
25. MoothaVKLepagePMillerKBunkenborgJReichM 2003 Identification of a gene causing human cytochrome c oxidase deficiency by integrative genomics. Proc Natl Acad Sci USA 100 605 610 doi:10.1073/pnas.242716699
26. MantheyGMMcEwenJE 1995 The product of the nuclear gene PET309 is required for translation of mature mRNA and stability or production of intron-containing RNAs derived from the mitochondrial COX1 locus of Saccharomyces cerevisiae. EMBO J 14 4031 4043
27. MantheyGMPrzybyla-ZawislakBDMcEwenJE 1998 The Saccharomyces cerevisiae Pet309 protein is embedded in the mitochondrial inner membrane. Eur J Biochem 255 156 161
28. SanchiricoMEFoxTDMasonTL 1998 Accumulation of mitochondrially synthesized Saccharomyces cerevisiae Cox2p and Cox3p depends on targeting information in untranslated portions of their mRNAs. EMBO J 17 5796 5804 doi:10.1093/emboj/17.19.5796
29. SachsAB 1993 Messenger RNA degradation in eukaryotes. Cell 74 413 421
30. TemperleyRJSenecaSHTonskaKBartnikEBindoffLA 2003 Investigation of a pathogenic mtDNA microdeletion reveals a translation-dependent deadenylation decay pathway in human mitochondria. Hum Mol Genet 12 2341 2348 doi:10.1093/hmg/ddg238
31. Chrzanowska-LightowlersZMPreissTLightowlersRN 1994 Inhibition of mitochondrial protein synthesis promotes increased stability of nuclear-encoded respiratory gene transcripts. J Biol Chem 269 27322 27328
32. MetodievMDLeskoNParkCBCamaraYShiY 2009 Methylation of 12S rRNA is necessary for in vivo stability of the small subunit of the mammalian mitochondrial ribosome. Cell Metab 9 386 397 doi:10.1016/j.cmet.2009.03.001
33. AltschulSFMaddenTLSchäfferAAZhangJZhangZ 1997 Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25 3389 3402
34. ChennaRSugawaraHKoikeTLopezRGibsonTJ 2003 Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res 31 3497 3500
35. TalaveraGCastresanaJ 2007 Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Syst Biol 56 564 577 doi:10.1080/10635150701472164
36. GuindonSDelsucFDufayardJ-FGascuelO 2009 Estimating maximum likelihood phylogenies with PhyML. Methods Mol Biol 537 113 137 doi:10.1007/978-1-59745-251-9_6
37. HusonDHRichterDCRauschCDezulianTFranzM 2007 Dendroscope: An interactive viewer for large phylogenetic trees. BMC Bioinformatics 8 460 doi:10.1186/1471-2105-8-460
38. CoxBEmiliA 2006 Tissue subcellular fractionation and protein extraction for use in mass-spectrometry-based proteomics. Nat Protoc 1 1872 1878 doi:10.1038/nprot.2006.273
39. GreeneJCWhitworthAJKuoIAndrewsLAFeanyMB 2003 Mitochondrial pathology and apoptotic muscle degeneration in Drosophila parkin mutants. Proc Natl Acad Sci USA 100 4078 4083 doi:10.1073/pnas.0737556100
40. FreyerCParkCBEkstrandMIShiYKhvorostovaJ 2010 Maintenance of respiratory chain function in mouse hearts with severely impaired mtDNA transcription. Nucleic Acids Res 38 6577 6588 doi:10.1093/nar/gkq527
41. Fernández-MorenoMAFarrCLKaguniLSGaresseR 2007 Drosophila melanogaster as a model system to study mitochondrial biology. Methods Mol Biol 372 33 49
42. WredenbergAWibomRWilhelmssonHGraffCWienerHH 2002 Increased mitochondrial mass in mitochondrial myopathy mice. Proc Natl Acad Sci USA 99 15066 15071 doi:10.1073/pnas.232591499
43. KimmichGARandlesJBrandJS 1975 Assay of picomole amounts of ATP, ADP, and AMP using the luciferase enzyme system. Anal Biochem 69 187 206
44. MourierADevinARigouletM 2010 Active proton leak in mitochondria: a new way to regulate substrate oxidation. Biochim Biophys Acta 1797 255 261 doi:10.1016/j.bbabio.2009.10.011
45. EnríquezJAPérez-MartosALópez-PérezMJMontoyaJ 1996 In organello RNA synthesis system from mammalian liver and brain. Meth Enzymol 264 50 57
46. CôtéCPoirierJBouletD 1989 Expression of the mammalian mitochondrial genome. Stability of mitochondrial translation products as a function of membrane potential. J Biol Chem 264 8487 8490
47. RobertiMBruniFLoguercio PolosaPManzariCGadaletaMN 2006 MTERF3, the most conserved member of the mTERF-family, is a modular factor involved in mitochondrial protein synthesis. Biochim Biophys Acta 1757 1199 1206 doi:10.1016/j.bbabio.2006.04.026
48. SasarmanFAntonickaHShoubridgeEA 2008 The A3243G tRNALeu(UUR) MELAS mutation causes amino acid misincorporation and a combined respiratory chain assembly defect partially suppressed by overexpression of EFTu and EFG2. Hum Mol Genet 17 3697 3707 doi:10.1093/hmg/ddn265
49. CouttetPFromont-RacineMSteelDPictetRGrangeT 1997 Messenger RNA deadenylylation precedes decapping in mammalian cells. Proc Natl Acad Sci USA 94 5628 5633
50. StewartJBBeckenbachAT 2009 Characterization of mature mitochondrial transcripts in Drosophila, and the implications for the tRNA punctuation model in arthropods. Gene 445 49 57 doi:10.1016/j.gene.2009.06.006
51. SchäggerHJagow vonG 1991 Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal Biochem 199 223 231
52. DekkerPJMüllerHRassowJPfannerN 1996 Characterization of the preprotein translocase of the outer mitochondrial membrane by blue native electrophoresis. Biol Chem 377 535 538
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
2011 Číslo 10
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