Molecular Genetics and Genomics

, Volume 274, Issue 4, pp 410–418

UPR-independent dithiothreitol stress-induced genes in Aspergillus niger

  • D. A. MacKenzie
  • T. Guillemette
  • H. Al-Sheikh
  • A. J. Watson
  • D. J. Jeenes
  • P. Wongwathanarat
  • N. S. Dunn-Coleman
  • N. van. Peij
  • D. B. Archer
Original Paper

DOI: 10.1007/s00438-005-0034-3

Cite this article as:
MacKenzie, D.A., Guillemette, T., Al-Sheikh, H. et al. Mol Genet Genomics (2005) 274: 410. doi:10.1007/s00438-005-0034-3

Abstract

A subtraction library was prepared from cultures of Aspergillus niger that had or had not been exposed to dithiothreitol (DTT), in order to identify genes involved in the unfolded protein response (UPR) or in the response to reductive stress. A large fraction of the clones in the library (40%) encoded two putative methyltransferases (MTs) whose function has yet to be determined. Other stress-responsive genes included a homologue of the Mn2+ -containing superoxide dismutase gene (sodB) and a number of genes predicted to code for products that function in protein turnover and in intra- and extracellular transport of molecules. Transcriptional microarray analysis was carried out with a group of 15 genes, comprising 11 from the cDNA library, two genes linked to the putative MT genes but not represented in the library, and two UPR control genes (bipA and pdiA). Eleven of the 15 genes were inducible with DTT. This was either reflected by the presence of transcripts in cells subjected to DTT stress compared to absence under control conditions, or by an induction ratio of between 1.4 and 8.0 in cases where transcripts were already detectable under control conditions. The MT genes were among the four most highly induced. None of the genes, apart from bipA and pdiA, showed significant induction in response to other stresses that are known to induce the UPR in fungi. We conclude that DTT alone does not provide for specific induction of UPR genes and that other stress conditions must also be examined.

Keywords

PCR subtractionReducing agentProtein secretionMethyltransferasesFilamentous fungi

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • D. A. MacKenzie
    • 1
  • T. Guillemette
    • 2
  • H. Al-Sheikh
    • 2
  • A. J. Watson
    • 2
    • 6
  • D. J. Jeenes
    • 1
  • P. Wongwathanarat
    • 3
  • N. S. Dunn-Coleman
    • 4
  • N. van. Peij
    • 5
  • D. B. Archer
    • 2
  1. 1.Department of Food Safety ScienceInstitute of Food ResearchColney, NorwichUK
  2. 2.School of BiologyUniversity of NottinghamNottinghamUK
  3. 3.Department of Biotechnology, Faculty of Science and TechnologyThammasat UniversityKhlong Luang, Pathum ThaniThailand
  4. 4.Genencor International Inc.Palo AltoUSA
  5. 5.Research and DevelopmentDSM Food SpecialtiesDelftThe Netherlands
  6. 6.School of BiologyUniversity of Newcastle-upon-TyneNewcastle-upon-TyneUK