Important progress has been made in the technology of optogenetic control of proteins in living cells

Recently, the team of Professor Yang Yi of the School of Pharmacy of East China University of Science and Technology published a paper in Nature Communications, describing an ultra-sensitive light-induced protein stability tag that can be used to regulate protein stability in living cells.

Live-cell protein optogenetic control. Photo courtesy of East China University of Science and Technology

Gene editing, transcriptional regulation, and RNA interference are widely used methods for protein manipulation in living cells that can be used to study the function of specific proteins in complex biological processes. As a flexible and powerful genome editing tool, the CRISPR-Cas system has been widely used in recent years. However, these techniques control proteins at the gene or mRNA level, and mRNA transcription and translation take time, resulting in significant delays in phenotypic control of these methods. Therefore, it is critical to develop tools for high-spatiotemporal resolution regulation of proteins directly at the protein level.

Due to its precise temporal and spatial resolution, light can meet most requirements for protein control switching. To date, scientists have designed a variety of light-induced degraders (LIDs) and successfully applied them to the precise optogenetic regulation of protein degradation in model organisms such as yeast, nematodes, zebrafish, and mammalian cells. However, these LIDs destabilize the target protein even in the non-induced state, thereby narrowing the dynamic regulatory range at the protein level. In addition, the degrador short peptides in LIDs can only function at the C-terminus of the protein, and LIDs must also fuse at the C-terminus of the target protein, which is not suitable for proteins that do not tolerate C-terminal fusion. Therefore, the development of light-sensitive protein degraders with excellent light-induced properties and better versatility remains urgent and challenging.

In this study, the research team developed the light-induced stable protein degradation tag SULI based on the photosensitive protein VVD. In dark conditions, the SULI tag is recognized and degraded by the cell’s degradation system; Under blue light irradiation, the SULI tag is very stable.

“The experimental results show that SUFI can be used to regulate the stability of a variety of reporter proteins. More importantly, SULI can be inserted into the N-terminus, C-terminus or even inside the protein of interest to regulate their stability. Dr. Mao Miaowei, the first author of the paper, said.

Further studies have shown that SULI is highly dependent on the degradation of the protein of interest, and can oscillatively regulate the abundance of the protein of interest in cells. The team also explored the molecular mechanism of SULI-mediated protein degradation and found that SULI is degraded through a non-lysine ubiquitination-dependent proteasome pathway, accompanied by aggregation and deaggregation processes. The researchers used microscopic imaging experiments to prove that SULI aggregates during its degradation, deaggregates with the assistance of a protein and enters the proteasome to complete the degradation process. The researchers then used SULI to regulate the stability of endogenous proteins in yeast cells, achieving precise control of the yeast cell cycle. In addition, SULI can also be used to regulate the stability of proteins in zebrafish, and achieve precise control of the development process of zebrafish through the regulation of the stability of a protein. This provides a powerful and convenient tool for studying protein metabolism and function in different cellular processes. (Source: China Science News, Zhang Shuanghu, Li Chenyang)

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