Isolation and characterisation of a novel stress-inducible PDI-family gene from Aspergillus niger.
Current strategies to improve the secretion of heterologous proteins in Aspergillus niger include the manipulation of chaperones and foldases specific to the endoplasmic reticulum (ER). A family of ER-specific proteins which share active-site homology wit protein disulfide isomerase (PDI) has been identified from other systems, many of which are inducible by agents which cause malfolding of proteins in the ER. Here we report identification of tigA from Aspergillus niger and erp38 from Neurospora crassa, two novel members of the PDI superfamily of proteins. TIGA and ERp38 show 66% identity at the amino acid level and are putative ER proteins. Both proteins show tandemly linked thiol-oxidoreductase domains followed by a functionally uncharacterised C-terminal domain. The most distal active site in TIGA is created by excision of a 66-bp intron. Although no Unfolded Protein Response elements can be seen in the tigA promoter, sequence homology has identified associated with protein trafficking (ERPTRE) in a gene encoding the related mammalian protein, ERp72, as well as a second motif conserved amongst the glucose-related protein family. Southern and dot blot analysis indicate that the tigA gene is present in single copy. Both the A. niger and N. crassa proteins show homology with a stress-inducible alfalfa, G1. Transcription of tigA is induced 2-3-fold after treatment with tunicamycin, an inhibitor of N-linked glycosylation. Strains overexpressing a heterologous protein show no increased tigA mRNA levels.
Jeenes
D J
DJ
Department of Genetics and Microbiology, Institute of Food Research, Colney, Norwich, UK. david.jeenes@bbsrc.ac.uk
Pfaller
R
R
Archer
D B
DB
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005656:Fungal Proteins; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D006360:Heat-Shock Proteins / Q000235:genetics*; D007535:Isomerases / Q000235:genetics*; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000201:enzymology / Q000235:genetics; D016366:Open Reading Frames; D019704:Protein Disulfide-Isomerases; D017510:Protein Folding; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D018076:DNA, Complementary; D005656:Fungal Proteins; D006360:Heat-Shock Proteins; C107551:TigA protein, Aspergillus niger; D007535:Isomerases; C107552:ERp38 protein, Neurospora crassa; D019704:Protein Disulfide-Isomerases