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Split invertase polypeptides form functional complexes in the yeast periplasm in vivo
O Schonberger1, C Knox, E Bibi
1Department of Molecular Biology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Summary
Split gene products can assemble into functional invertase enzyme within the yeast secretory pathway. This protein complex forms in the endoplasmic reticulum and facilitates growth on sucrose.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Protein assembly from fragments in vivo has been shown for some proteins.
- The function of split gene products within eukaryotic secretory systems remains largely unexplored.
Purpose of the Study:
- To investigate the functionality of split gene products within the eukaryotic secretory pathway.
- To determine if split invertase fragments can reassemble into an active enzyme in Saccharomyces cerevisiae.
Main Methods:
- Utilized Saccharomyces cerevisiae strains engineered to produce invertase.
- Expressed independently translated and translocated overlapping fragments of invertase.
- Assessed enzymatic activity and biological function of the reassembled invertase complex.
Main Results:
- Invertase fragments with signal peptides were translocated, glycosylated, and trafficked through the secretory pathway.
- Simultaneous expression of overlapping fragments resulted in an active invertase complex, while individual fragments were inactive.
- The assembled invertase complex was biologically functional, enabling yeast growth on sucrose.
Conclusions:
- Split gene products can function within the eukaryotic secretory pathway.
- Protein assembly and functional complex formation can occur within the endoplasmic reticulum.
- This study provides insights into protein folding, assembly, and trafficking in the secretory pathway.