Eisosome disruption by noncoding RNA deletion increases protein secretion in yeast

Area of Science:

• Molecular Biology
• Biotechnology
• Yeast Genetics

Background:

• Noncoding RNAs (ncRNAs) play crucial roles in regulating gene expression.
• Efficient protein secretion in yeast is vital for biotechnology and biopharmaceutical production.
• Eisosomes, plasma membrane invaginations, are involved in cellular organization and transport.

Purpose of the Study:

• To investigate the impact of ncRNA deletions on endogenous invertase secretion in yeast.
• To identify specific ncRNAs that, when deleted, enhance protein secretion.
• To explore the potential of eisosome disruption as a strategy for improving protein secretion.

Main Methods:

• Large-scale screening of ncRNA deletion strains (SUTs, CUTs, tRNAs, snRNAs) for improved invertase secretion.
• Quantification of adjacent gene transcription in ncRNA deletion strains.
• Analysis of protein secretion (invertase, GFP, monoclonal antibody peptide) in knockout strains and upon eisosome disruption.

Main Results:

• Deletion of SUT418, SUT390, and SUT125 significantly improved invertase secretion.
• PIL1 gene down-regulation in the SUT125 deletion strain led to increased invertase secretion, further enhanced by combined deletions of eisosome components.
• Eisosome disruption increased secretion of various proteins, including heterologous proteins and a monoclonal antibody peptide, linked to sphingolipid metabolism alterations.

Conclusions:

• Specific ncRNAs, such as SUT125, can be deleted to enhance protein secretion.
• Disruption of eisosomes, particularly through PIL1 deletion, represents a novel pathway to improve both endogenous and heterologous protein secretion in yeast.
• This strategy holds potential for enhancing biopharmaceutical production using engineered yeast systems.