Increased production of secreted proteins by recombinant eukaryotic cells

Abstract

Described herein are methods for increasing the amount of protein secreted by a cell. In one case, a cell is provided which contains a heterologous nucleic acid encoding a protein having unfolded protein response modulating activity and a heterologous nucleic acid encoding a protein of interest to be secreted. In one case, the protein having unfolded protein response modulating activity is selected from the proteins selected from the group consisting of HAC1, PTC2 and IRE1. The protein of interest can be any secreted protein such as a therapeutic or an industrial enzyme. For example the protein can be selected from the group consisting of lipase, cellulase, endo-glucosidase H, protease, carbohydrase, reductase, oxidase, isomerase, transferase, kinase, phosphatase, alpha-amylase, glucoamylase, lignocellulose hemicellulase, pectinase and ligninase.

What is claimed is: 
     
         1 - 55 . (canceled) 
     
     
         56 . An isolated nucleic acid encoding a PTC2 protein wherein said PTC2 protein modulates unfolded protein response and wherein said PTC2 has at least 70% similarity to an amino acid sequence of  FIG. 24  or  FIG. 25 . 
     
     
         57 . The nucleic acid of  claim 56  isolated from  Trichoderma reesei.   
     
     
         58 . The nucleic acid of  claim 56  isolated from  Aspergillus nidulans.    
     
     
         59 . The nucleic acid of  claim 56  isolated from  Aspergillus niger.   
     
     
         60 . The nucleic acid of  claim 56  wherein said PTC2 protein has an amino acid sequence as set forth in  FIG. 24 . 
     
     
         61 . The nucleic acid of  claim 56  wherein said PTC2 protein has an amino acid sequence as set forth in  FIG. 25 . 
     
     
         62 . The nucleic acid of  claim 56  wherein said nucleic acid comprises a coding nucleic acid sequence as set forth in  FIG. 24 . 
     
     
         63 . The nucleic acid of  claim 56  wherein said nucleic acid consists essentially of a coding nucleic acid sequence as set forth in  FIG. 24 . 
     
     
         64 . The nucleic acid of  claim 56  wherein said nucleic acid comprises a coding nucleic acid sequence as set forth in  FIG. 25 . 
     
     
         65 . A protein encoded by the nucleic acid of  claim 56 . 
     
     
         66 . A protein having unfolded protein response modulating activity and having greater than 70% similarity to the amino acid sequence of  FIG. 24  or  FIG. 25 . 
     
     
         67 . A protein having an amino acid sequence as set forth in  FIG. 24  or  FIG. 25 . 
     
     
         68 . A nucleic acid encoding a IRE1 protein having unfolded protein response modulating activity and having at least 60% to an amino acid sequence as shown in  FIG. 26  or  FIG. 27 . 
     
     
         69 . The nucleic acid of  claim 68  wherein said IRE1 protein has an amino acid sequence as shown in  FIG. 26  or  FIG. 27 . 
     
     
         70 . The nucleic acid of  claim 68  wherein said nucleic acid is isolated from  Trichoderma reesei.   
     
     
         71 . The nucleic acid of  claim 68  wherein said nucleic acid is isolated from  Aspergillus nidulans.   
     
     
         72 . The nucleic acid of  claim 68  wherein said nucleic acid is isolated from  Aspergillus niger.   
     
     
         73 . The nucleic acid of  claim 68  wherein said IRE1 protein has an amino acid sequence as set forth in  FIG. 26 . 
     
     
         74 . The nucleic acid of  claim 68  wherein said IRE1 protein has an amino acid sequence as set forth in  FIG. 27 . 
     
     
         75 . The nucleic acid of  claim 68  wherein said nucleic acid comprises a coding nucleic acid sequence as set forth in  FIG. 26 . 
     
     
         76 . The nucleic acid of  claim 68  wherein said nucleic acid consists essentially of a coding nucleic acid sequence as set forth in  FIG. 26 . 
     
     
         77 . The nucleic acid of  claim 68  wherein said nucleic acid comprises a coding nucleic acid sequence as set forth in  FIG. 27 . 
     
     
         78 . The nucleic acid of  claim 68  wherein said nucleic acid consists essentially of a coding nucleic acid sequence as set forth in  FIG. 27 . 
     
     
         79 . A protein encoded by the nucleic acid of  claim 68 . 
     
     
         80 . A protein having unfolded protein response inducing activity and having greater than 70% similarity to the amino acid sequence of  FIG. 26  or  FIG. 27 . 
     
     
         81 . The protein of  claim 80  wherein said protein has constitutive unfolded protein response inducing activity. 
     
     
         82 . A protein having an amino acid sequence as set forth in  FIG. 26  or  FIG. 27 . 
     
     
         83 - 86 . (canceled)