Isolation and characterisation of a calnexin homologue, clxA, from Aspergillus niger.
We describe the isolation of a gene (clxA) encoding calnexin from laboratory and industrial strains of Aspergillus niger. Calnexin is a chaperone, which specifically recognises monoglucosylated glycoproteins in the endoplasmic reticulum, and is thus an essential component of the process that assesses the folded state of nascent secreted glycoproteins. Manipulation of chaperones has previously been adopted in attempts to overcome some of the problems associated with the secretion of heterologous proteins from filamentous fungi. The A. niger clxA gene encodes a 562-residue protein with strong homology to the calnexin of Schizosaccharomyces pombe. The clxAgene product complements a S. pombe cnx1 mutant. Motifs associated with genes controlled via the Unfolded Protein Response (UPR) were identified by sequence homology in the promoter of clxA. Steady-state levels of clxA mRNA were elevated in a strain expressing bovine prochymosin fused to the catalytic domain of glucoamylase. The ORF is punctuated by four introns, and contains two sets of four repeated peptide motifs that are characteristic of the calnexin family, together with a putative membrane-spanning domain. Deletion studies indicate that clxA is not an essential gene in A. niger.
Wang
H
H
Genencor International Inc., Palo Alto, CA 94304, USA.
Entwistle
J
J
Morlon
E
E
Archer
D B
DB
Peberdy
J F
JF
Ward
M
M
Jeenes
D J
DJ
D000818:Animals; D001234:Aspergillus niger / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D037281:Calnexin / Q000235:genetics* / Q000378:metabolism; D002417:Cattle; D012085:Chymosin / Q000096:biosynthesis / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004792:Enzyme Precursors / Q000096:biosynthesis / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D015870:Gene Expression; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D017386:Sequence Homology, Amino Acid
C094757:Cnx1 protein, S pombe; D004271:DNA, Fungal; D004792:Enzyme Precursors; D005656:Fungal Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D037281:Calnexin; D005087:Glucan 1,4-alpha-Glucosidase; C032443:prorennin; D012085:Chymosin