UPR-independent dithiothreitol stress-induced genes in Aspergillus niger.
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.
MacKenzie
D A
DA
Department of Food Safety Science, Institute of Food Research, Norwich Research Park, Colney, Norwich, NR4 7UA, UK. donald.mackenzie@bbsrc.ac.uk
Guillemette
T
T
Al-Sheikh
H
H
Watson
A J
AJ
Jeenes
D J
DJ
Wongwathanarat
P
P
Dunn-Coleman
N S
NS
van Peij
N
N
Archer
D B
DB
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000235:genetics* / Q000378:metabolism*; D018076:DNA, Complementary / Q000378:metabolism; D004229:Dithiothreitol / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress; D010957:Plasmids / Q000378:metabolism; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D018076:DNA, Complementary; D005656:Fungal Proteins; D004229:Dithiothreitol