Skip to main content
Log in

Different physiological relevance of yeast THO/TREX subunits in gene expression and genome integrity

  • Original Paper
  • Published:
Molecular Genetics and Genomics Aims and scope Submit manuscript

Abstract

THO/TREX is a conserved nuclear complex that functions in mRNP biogenesis and plays a role in preventing the transcription-associated genetic instability. THO is composed of Tho2, Hpr1, Mft1 and Thp2 subunits, which associate with the Sub2-Yra1 export factors and Tex1 to form the TREX complex. To compare the functional relevance of the different THO/TREX subunits, we determined the effect of their null mutations on mRNA accumulation and recombination. Unexpectedly, we noticed that a full deletion of HPR1, hpr1ΔK, conferred stronger hyper-recombination phenotype and gene expression defects than did hpr1ΔH, the allele encoding a C-terminal truncated protein which was used in most previous studies. We show that tho2Δ and, to a lesser extent, hpr1ΔK are the THO mutations with the highest impact on all phenotypes, and that sub2Δ shows a similar transcription-dependent hyper-recombination phenotype and in vivo transcription impairment as hpr1ΔK and tho2Δ. Recombination and transcription analyses indicate that THO/TREX mutants share a moderate but significant effect on gene conversion and ectopic recombination, as well as transcription impairment of even short and low GC-content genes. Our data provide new information on the relevance of these proteins in mRNP biogenesis and in the maintenance of genomic integrity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Aguilera A (2002) The connection between transcription and genomic instability. EMBO J 21:195–201

    Article  PubMed  CAS  Google Scholar 

  • Aguilera A (2005) Cotranscriptional mRNP assembly: from the DNA to the nuclear pore. Curr Opin Cell Biol 17:242–250

    Article  PubMed  CAS  Google Scholar 

  • Aguilera A, Klein HL (1988) Genetic control of intrachromosomal recombination in Saccharomyces cerevisiae.I.Isolation and genetic characterization of hyper-recombination mutations. Genetics 119:779–790

    PubMed  CAS  Google Scholar 

  • Aguilera A, Klein HL (1990) HPR1, a novel yeast gene that prevents intrachromosomal excision recombination, shows carboxy-terminal homology to the Saccharomyces cerevisiae TOP1 gene. Mol Cell Biol 10:1439–1451

    PubMed  CAS  Google Scholar 

  • Burckin T et al. (2005) Exploring functional relationships between components of the gene expression machinery. Nat Struct Mol Biol 12:175–182

    Article  PubMed  CAS  Google Scholar 

  • Chang M, French-Cornay D, Fan HY, Klein H, Denis CL, Jaehning JA (1999) A complex containing RNA polymerase II, Paf1p, Cdc73p, Hpr1p, and Ccr4p plays a role in protein kinase C signaling. Mol Cell Biol 19:1056–1067

    PubMed  CAS  Google Scholar 

  • Chávez S et al. (2000) A protein complex containing Tho2, Hpr1, Mft1 and a novel protein, Thp2, connects transcription elongation with mitotic recombination in Saccharomyces cerevisiae. EMBO J 19:5824–5834

    Article  PubMed  Google Scholar 

  • Chávez S, García-Rubio M, Prado F, Aguilera A (2001) Hpr1 is preferentially required for transcription of either long or G + C-rich DNA sequences in Saccharomyces cerevisiae. Mol Cell Biol 21:7054–7064

    Article  PubMed  Google Scholar 

  • Fan HY, Merker RJ, Klein HL (2001) High-copy-number expression of Sub2p, a member of the RNA helicase superfamily, suppresses hpr1-mediated genomic instability. Mol Cell Biol 21:5459–5470

    Article  PubMed  CAS  Google Scholar 

  • Fischer T et al. (2002) The mRNA export machinery requires the novel Sac3p-Thp1p complex to dock at the nucleoplasmic entrance of the nuclear pores. EMBO J 21:5843–5852

    Article  PubMed  CAS  Google Scholar 

  • Freedman JA, Jinks-Robertson S (2004) Effects of mismatch repair and Hpr1 on transcription-stimulated mitotic recombination in the yeast Saccharomyces cerevisiae. DNA Repair (Amst) 3:1437–1446

    Article  CAS  Google Scholar 

  • Gaillard H, Wellinger RE, Aguilera A (2007) A new connection of mRNP biogenesis and export with transcription-coupled repair. Nucleic Acids Res 35:3893–3906

    Article  PubMed  CAS  Google Scholar 

  • Gallardo M, Luna R, Erdjument-Bromage H, Tempst P, Aguilera A (2003) Nab2p and the Thp1p-Sac3p complex functionally interact at the interface between transcription and mRNA metabolism. J Biol Chem 278:24225–24232

    Article  PubMed  CAS  Google Scholar 

  • Gari E, Piedrafita L, Aldea M, Herrero E (1997) A set of vectors with a tetracycline-regulatable promoter system for modulated gene expression in Saccharomyces cerevisiae. Yeast 13:837–848

    Article  PubMed  CAS  Google Scholar 

  • Huertas P, Aguilera A (2003) Cotranscriptionally formed DNA:RNA hybrids mediate transcription elongation impairment and transcription-associated recombination. Mol Cell 12:711–721

    Article  PubMed  CAS  Google Scholar 

  • Huertas P, García-Rubio ML, Wellinger RE, Luna R, Aguilera A (2006) An hpr1 point mutation that impairs transcription and mRNP biogenesis without increasing recombination. Mol Cell Biol 26:7451–7465

    Article  PubMed  CAS  Google Scholar 

  • Jimeno S, Rondón AG, Luna R, Aguilera A (2002) The yeast THO complex and mRNA export factors link RNA metabolism with transcription and genome instability. EMBO J 21:3526–3535

    Article  PubMed  CAS  Google Scholar 

  • Kim M, Ahn SH, Krogan NJ, Greenblatt JF, Buratowski S (2004) Transitions in RNA polymerase II elongation complexes at the 3′ ends of genes. EMBO J 23:354–364

    Article  PubMed  CAS  Google Scholar 

  • Libri D, Dower K, Boulay J, Thomsen R, Rosbash M, Jensen TH (2002) Interactions between mRNA export commitment, 3′-end quality control, and nuclear degradation. Mol Cell Biol 22:8254–8266

    Article  PubMed  CAS  Google Scholar 

  • Libri D, Graziani N, Saguez C, Boulay J (2001) Multiple roles for the yeast SUB2/yUAP56 gene in splicing. Genes Dev 15:36–41

    Article  PubMed  CAS  Google Scholar 

  • Luna R, Jimeno S, Marin M, Huertas P, García-Rubio M, Aguilera A (2005) Interdependence between transcription and mRNP processing and export, and its impact on genetic stability. Mol Cell 18:711–722

    Article  PubMed  CAS  Google Scholar 

  • Mason PB, Struhl K (2005) Distinction and relationship between elongation rate and processivity of RNA polymerase II in vivo. Mol Cell 17:831–840

    Article  PubMed  CAS  Google Scholar 

  • Morillo-Huesca M, Vanti M, Chávez S (2006) A simple in vivo assay for measuring the efficiency of gene length-dependent processes in yeast mRNA biogenesis. FEBS J 273:756–769

    Article  PubMed  CAS  Google Scholar 

  • Piruat JI, Aguilera A (1998) A novel yeast gene, THO2, is involved in RNA pol II transcription and provides new evidence for transcriptional elongation-associated recombination. EMBO J 17:4859–4872

    Article  PubMed  CAS  Google Scholar 

  • Prado F, Piruat JI, Aguilera A (1997) Recombination between DNA repeats in yeast hpr1delta cells is linked to transcription elongation. EMBO J 16:2826–2835

    Article  PubMed  CAS  Google Scholar 

  • Rehwinkel J et al. (2004) Genome-wide analysis of mRNAs regulated by the THO complex in Drosophila melanogaster. Nat Struct Mol Biol 11:558–566

    Article  PubMed  CAS  Google Scholar 

  • Rondón AG, García-Rubio M, Gonzalez-Barrera S, Aguilera A (2003a) Molecular evidence for a positive role of Spt4 in transcription elongation. EMBO J 22:612–620

    Article  PubMed  Google Scholar 

  • Rondón AG, Jimeno S, García-Rubio M, Aguilera A (2003b) Molecular evidence that the eukaryotic THO/TREX complex is required for efficient transcription elongation. J Biol Chem 278:39037–39043

    Article  PubMed  Google Scholar 

  • Santos-Rosa H, Aguilera A (1994) Increase in incidence of chromosome instability and non-conservative recombination between repeats in Saccharomyces cerevisiae hpr1 delta strains. Mol Gen Genet 245:224–236

    Article  PubMed  CAS  Google Scholar 

  • Strasser K et al. (2002) TREX is a conserved complex coupling transcription with messenger RNA export. Nature 417:304–308

    Article  PubMed  Google Scholar 

  • Vinciguerra P, Stutz F (2004) mRNA export: an assembly line from genes to nuclear pores. Curr Opin Cell Biol 16:285–292

    Article  PubMed  CAS  Google Scholar 

  • Voynov V, Verstrepen KJ, Jansen A, Runner VM, Buratowski S, Fink GR (2006) Genes with internal repeats require the THO complex for transcription. Proc Natl Acad Sci USA 103:14423–14428

    Article  PubMed  CAS  Google Scholar 

  • Wach A, Brachat A, Alberti-Segui C, Rebischung C, Philippsen P (1997) Heterologous HIS3 marker and GFP reporter modules for PCR-targeting in Saccharomyces cerevisiae. Yeast 13:1065–1075

    Article  PubMed  CAS  Google Scholar 

  • Wang X, Chang Y, Li Y, Zhang X, Goodrich DW (2006) Thoc1/Hpr1/p84 is essential for early embryonic development in the mouse. Mol Cell Biol 26:4362–4367

    Article  PubMed  CAS  Google Scholar 

  • Wellinger RE, Prado F, Aguilera A (2006) Replication fork progression is impaired by transcription in hyperrecombinant yeast cells lacking a functional THO complex. Mol Cell Biol 26:3327–3334

    Article  PubMed  CAS  Google Scholar 

  • Zenklusen D, Vinciguerra P, Wyss JC, Stutz F (2002) Stable mRNP formation and export require cotranscriptional recruitment of the mRNA export factors Yra1p and Sub2p by Hpr1p. Mol Cell Biol 22:8241–8253

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank H. Gaillard for critical reading of the manuscript, and D. Haun for style supervision. This work was supported by the Ministry of Science and Education of Spain (grants SAF2003–00204 and BMC2006–05260) and Junta de Andalucía (CVI-102).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Rosa Luna.

Additional information

Communicated by S. Hohmann.

Rights and permissions

Reprints and permissions

About this article

Cite this article

García-Rubio, M., Chávez, S., Huertas, P. et al. Different physiological relevance of yeast THO/TREX subunits in gene expression and genome integrity. Mol Genet Genomics 279, 123–132 (2008). https://doi.org/10.1007/s00438-007-0301-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00438-007-0301-6

Keywords

Navigation