3′ Untranslated Regions Mediate Transcriptional Interference between Convergent Genes Both Locally and Ectopically in
Paired sense and antisense (S/AS) genes located in cis represent a structural feature common to the genomes of both prokaryotes and eukaryotes, and produce partially complementary transcripts. We used published genome and transcriptome sequence data and found that over 20% of genes (645 pairs) in the budding yeast Saccharomyces cerevisiae genome are arranged in convergent pairs with overlapping 3′-UTRs. Using published microarray transcriptome data from the standard laboratory strain of S. cerevisiae, our analysis revealed that expression levels of convergent pairs are significantly negatively correlated across a broad range of environments. This implies an important role for convergent genes in the regulation of gene expression, which may compensate for the absence of RNA-dependent mechanisms such as micro RNAs in budding yeast. We selected four representative convergent gene pairs and used expression assays in wild type yeast and its genetically modified strains to explore the underlying patterns of gene expression. Results showed that convergent genes are reciprocally regulated in yeast populations and in single cells, whereby an increase in expression of one gene produces a decrease in the expression of the other, and vice-versa. Time course analysis of the cell cycle illustrated the functional significance of this relationship for the three pairs with relevant functional roles. Furthermore, a series of genetic modifications revealed that the 3′-UTR sequence plays an essential causal role in mediating transcriptional interference, which requires neither the sequence of the open reading frame nor the translation of fully functional proteins. More importantly, transcriptional interference persisted even when one of the convergent genes was expressed ectopically (in trans) and therefore does not depend on the cis arrangement of convergent genes; we conclude that the mechanism of transcriptional interference cannot be explained by the transcriptional collision model, which postulates a clash between simultaneous transcriptional processes occurring on opposite DNA strands.
Vyšlo v časopise:
3′ Untranslated Regions Mediate Transcriptional Interference between Convergent Genes Both Locally and Ectopically in. PLoS Genet 10(1): e32767. doi:10.1371/journal.pgen.1004021
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.pgen.1004021
Souhrn
Paired sense and antisense (S/AS) genes located in cis represent a structural feature common to the genomes of both prokaryotes and eukaryotes, and produce partially complementary transcripts. We used published genome and transcriptome sequence data and found that over 20% of genes (645 pairs) in the budding yeast Saccharomyces cerevisiae genome are arranged in convergent pairs with overlapping 3′-UTRs. Using published microarray transcriptome data from the standard laboratory strain of S. cerevisiae, our analysis revealed that expression levels of convergent pairs are significantly negatively correlated across a broad range of environments. This implies an important role for convergent genes in the regulation of gene expression, which may compensate for the absence of RNA-dependent mechanisms such as micro RNAs in budding yeast. We selected four representative convergent gene pairs and used expression assays in wild type yeast and its genetically modified strains to explore the underlying patterns of gene expression. Results showed that convergent genes are reciprocally regulated in yeast populations and in single cells, whereby an increase in expression of one gene produces a decrease in the expression of the other, and vice-versa. Time course analysis of the cell cycle illustrated the functional significance of this relationship for the three pairs with relevant functional roles. Furthermore, a series of genetic modifications revealed that the 3′-UTR sequence plays an essential causal role in mediating transcriptional interference, which requires neither the sequence of the open reading frame nor the translation of fully functional proteins. More importantly, transcriptional interference persisted even when one of the convergent genes was expressed ectopically (in trans) and therefore does not depend on the cis arrangement of convergent genes; we conclude that the mechanism of transcriptional interference cannot be explained by the transcriptional collision model, which postulates a clash between simultaneous transcriptional processes occurring on opposite DNA strands.
Zdroje
1. LavorgnaG, DaharyD, LehnerB, SorekR, SandersonCM, et al. (2004) In search of antisense. Trends Biochem Sci 29: 88–94.
2. GuellM, van NoortV, YusE, ChenWH, Leigh-BellJ, et al. (2009) Transcriptome complexity in a genome-reduced bacterium. Science 326: 1268–1271.
3. ZhangY, LiuXS, LiuQR, WeiL (2006) Genome-wide in silico identification and analysis of cis natural antisense transcripts (cis-NATs) in ten species. Nucleic Acids Res 34: 3465–3475.
4. ChenN, SteinLD (2006) Conservation and functional significance of gene topology in the genome of Caenorhabditis elegans. Genome Res 16: 606–617.
5. KromN, RamakrishnaW (2008) Comparative analysis of divergent and convergent gene pairs and their expression patterns in rice, Arabidopsis, and populus. Plant Physiol 147: 1763–1773.
6. HeY, VogelsteinB, VelculescuVE, PapadopoulosN, KinzlerKW (2008) The antisense transcriptomes of human cells. Science 322: 1855–1857.
7. KatayamaS, TomaruY, KasukawaT, WakiK, NakanishiM, et al. (2005) Antisense transcription in the mammalian transcriptome. Science 309: 1564–1566.
8. NishizawaM, KomaiT, KatouY, ShirahigeK, ItoT, et al. (2008) Nutrient-regulated antisense and intragenic RNAs modulate a signal transduction pathway in yeast. PLoS Biol 6: 2817–2830.
9. HongayCF, GrisafiPL, GalitskiT, FinkGR (2006) Antisense transcription controls cell fate in Saccharomyces cerevisiae. Cell 127: 735–745.
10. CamblongJ, IglesiasN, FickentscherC, DieppoisG, StutzF (2007) Antisense RNA stabilization induces transcriptional gene silencing via histone deacetylation in S. cerevisiae. Cell 131: 706–717.
11. CamblongJ, BeyrouthyN, GuffantiE, SchlaepferG, SteinmetzLM, et al. (2009) Trans-acting antisense RNAs mediate transcriptional gene cosuppression in S. cerevisiae. Genes Dev 23: 1434–1445.
12. GullerovaM, ProudfootNJ (2008) Cohesin complex promotes transcriptional termination between convergent genes in S. pombe. Cell 132: 983–995.
13. PrescottEM, ProudfootNJ (2002) Transcriptional collision between convergent genes in budding yeast. Proc Natl Acad Sci USA 99: 8796–8801.
14. ShearwinKE, CallenBP, EganJB (2005) Transcriptional interference-a crash course. Trends Genet 21: 339–345.
15. FaghihiMA, WahlestedtC (2009) Regulatory roles of natural antisense transcripts. Nat Rev Mol Cell Biol 10: 637–643.
16. CramptonN, BonassWA, KirkhamJ, RivettiC, ThomsonNH (2006) Collision events between RNA polymerases in convergent transcription studied by atomic force microscopy. Nucleic Acids Res 34: 5416–5425.
17. NagalakshmiU, WangZ, WaernK, ShouC, RahaD, et al. (2008) The transcriptional landscape of the yeast genome defined by RNA sequencing. Science 320: 1344–1349.
18. ChurchmanLS, WeissmanJS (2011) Nascent transcript sequencing visualizes transcription at nucleotide resolution. Nature 469(20): 368–375.
19. ParkhomchukD, BorodinaT, AmstislavskiyV, BanaruM, HallenL, et al. (2009) Transcriptome analysis by strand-specific sequencing of complementary DNA. Nucleic Acids Res 37: e123.
20. HuXH, WangMH, TanT, LiJR, YangH, et al. (2007) Genetic dissection of ethanol tolerance in the budding yeast Saccharomyces cerevisiae. Genetics 175: 1479–1487.
21. WilsonWA, RoachPJ (2002) Nutrient-regulated protein kinases in budding yeast. Cell 111: 155–158.
22. Daignan-FornierB, FinkGR (1992) Coregulation of purine and histidine biosynthesis by the transcriptional activators BAS1 and BAS2. Proc Natl Acad Sci USA 89: 6746–6750.
23. StruhlK (1995) Yeast transcriptional regulatory mechanisms. Annu Rev Genetics 29: 651–674.
24. GoeffeauA, BarrellBG, BusseyH, DavisRW, DujonB, et al. (1996) Life with 6000 genes. Science 274: 546–567.
25. GullerovaM, MoazedD, ProudfootNJ (2011) Autoregulation of convergent RNAi genes in fission yeast. Genes Dev 25: 556–568.
26. DrinnenbergIA, WeinbergDE, XieKT, MowerJP, WolfeKH, et al. (2009) RNAi in budding yeast. Science 326: 544–550.
27. GrewalSI, JiaS (2007) Heterochromatin revisited. Nat Rev Genet 8: 35–46.
28. KitazonoAA, TobeBT, KaltonH, DiamantN, KronSJ (2002) Marker-fusion PCR for one-step mutagenesis of essential genes in yeast. Yeast 19: 141–149.
29. McNabbDS, ReedR, MarciniakRA (2005) Dual luciferase assay system for rapid assessment of gene expression in Saccharomyces cerevisiae. Eukaryot Cell 4: 1539–1549.
30. SchmittME, BrownTA, TrumpowerBL (1990) A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res 18: 3091–3092.
31. Del AguilaEM, DutraMB, SilvaJT, PaschoalinVM (2005) Comparing protocols for preparation of DNA-free total yeast RNA suitable for RT-PCR. BMC Mol Biol 6: 9.
32. YazawaH, IwahashiH, KamisakaY, KimuraK, UemuraH (2009) Production of polyunsaturated fatty acids in yeast Saccharomyces cerevisiae and its relation to alkaline pH tolerance. Yeast 26: 167–184.
33. LivakKJ, SchmittgenTD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25: 402–408.
34. EmmerthS, SchoberH, GaidatzisD, RoloffT, JacobeitK, et al. (2010) Nuclear retention of fission yeast dicer is a prerequisite for RNAi-mediated heterochromatin assembly. Dev Cell 18: 102–113.
35. SoutoG, GiacomettiR, SilbersteinS, GiassonL, CantoreML, et al. (2006) Expression of TPK1 and TPK2 genes encoding PKA catalytic subunits during growth and morphogenesis in Candida albicans. Yeast 23: 591–603.
36. EstrellaLA, WilkinsonMF, GonzalezCI (2009) The shuttling protein Npl3 promotes translation termination accuracy in Saccharomyces cerevisiae. J Mol Biol 394: 410–422.
37. FangF, SalmonK, ShenMW, AelingKA, ItoE, et al. (2011) A vector set for systematic metabolic engineering in Saccharomyces cerevisiae. Yeast 28: 123–136.
38. StowersCC, BoczkoEM (2007) Reliable cell disruption in yeast. Yeast 24: 533–541.
39. SpellmanPT, SherlockG, ZhangMQ, IyerVR, AndersK, et al. (1998) Comprehensive identification of cell cycle-regulated genes of the yeast Saccharomyces cerevisiae by microarray hybridization. Mol Biol Cell 9: 3273–3297.
40. FujitaM, HoriY, ShirahigeK, TsurimotoT, YoshikawaH, et al. (1998) Cell cycle dependent topological changes of chromosomal replication origins in Saccharomyces cerevisiae. Genes Cells 3: 737–749.
41. TangF, BarbacioruC, NordmanE, LiB, XuN, et al. (2010) RNA-Seq analysis to capture the transcriptome landscape of a single cell. Nat Protoc 5: 516–535.
42. IrizarryRA, HobbsB, CollinF, Beazer-BarclayYD, AntonellisKJ, et al. (2003) Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4: 249–264.
Štítky
Genetika Reprodukčná medicínaČlánok vyšiel v časopise
PLOS Genetics
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