Engineering degrons of yeast ornithine decarboxylase as vehicles for efficient targeted protein degradation

Joshi, Rushikesh G.; Kulkarni, Swapnali; Prabha, C. Ratna

doi:10.1016/j.bbagen.2015.09.003

Cited by 7 publications

(1 citation statement)

References 47 publications

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“…They originate from investigations of yeast or murine ornithine decarboxylase (ODC) degradation by the proteasome. Murine ODC carries a degron at the C-terminus that is activated upon antizyme binding to ODC; induction of protein degradation has been achieved in mammalian cell culture by fusing ODC to a target and manipulating antizyme levels. , Similarly, yeast ODC carries a degron at the N-terminus that is activated by the expression of yeast antizyme upon addition of a polyamine, e.g., putrescine, spermidine, or spermine. − However, usage of these conditional degrons requires the fusion of ODC to the target; ODC has a size of about 50 kDa, forms a dimer, and influences polyamine levels if the active enzyme is used.…”

Section: Conditional Control Of Protein Degradationmentioning

confidence: 99%

Synthetic Control of Protein Degradation during Cell Proliferation and Developmental Processes

et al. 2019

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Synthetic tools for the control of protein function are valuable for biomedical research to characterize cellular functions of essential proteins or if a rapid switch in protein activity is necessary. The ability to tune protein activity precisely opens another level of investigations that is not available with gene deletion mutants. Control of protein stability is a versatile approach to influence the activity of a target protein by its cellular abundance. Diverse strategies have been developed to achieve efficient proteolysis using external inducers or differentiation-coupled signals. The latter is especially important for the inactivation of a protein during a developmental process. Recently, several approaches to achieve this have been engineered. In this article, we present current synthetic tools for regulation of protein stability that allow fine-tuning of protein abundance, their advantages and disadvantages with an emphasis on methods applicable in the context of cell differentiation and development. We give an outlook toward future developments and discuss main applications of these tools.

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Section: Conditional Control Of Protein Degradationmentioning

confidence: 99%

Synthetic Control of Protein Degradation during Cell Proliferation and Developmental Processes

et al. 2019

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Degrons of yeast and mammalian ornithine decarboxylase enzymes make potent combination for regulated targeted protein degradation

Joshi

Prabha

2016

Appl Microbiol Biotechnol

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Elevation of polyamine levels in eukaryotes leads to rapid degradation of ornithine decarboxylase (ODC), the first enzyme of polyamine biosynthesis pathway. ODC in yeast (yODC) has two domains, the Nα/β domain consisting of α/β barrel domain (α/β) preceded by an overhang of 50 residues at its N-terminus (N50) and β sheet domain at its C-terminus. Two degradation determinant signals or degrons in yODC sequence, namely the N50 and the antizyme-binding element (AzBE) housed in the α/β domain, are responsible for its degradation by proteasomes. Antizyme (Az) induced under polyamine excess binds to AzBE and delivers ODC to proteasome, while the N50 threads the protein into proteasome. It was previously reported by us that the peptide Nα/β of yODC acts as an independent transplantable degron, whose action can be modulated with the help of antizyme by varying polyamine levels. Mammalian ODC (mODC), in spite of its 40% sequence homology with yODC, is devoid of N50 of yODC and instead sports a C-terminal tail of 37 residues (CmODC). CmODC acts as an independent transplantable degron with no equivalent in yODC. The present study investigates the merits of employing the two degrons Nα/β and CmODC together for targeted protein degradation by expressing them in a chimeric fusion with green fluorescent protein (GFP). Our results establish that under the regulation of antizyme, the signals Nα/β and CmODC acting together enhance degradation better than either degron in isolation. The combination of Nα/β and CmODC can be employed to study the function of novel proteins through their rapid removal.

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Structural and Functional Characterisation of the Domains of Ubiquitin-Activating Enzyme (E1) of Saccharomyces cerevisiae

Panchamia

Raimalani

Prashar

et al. 2020

Cell Biochem Biophys

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Engineering degrons of yeast ornithine decarboxylase as vehicles for efficient targeted protein degradation

Cited by 7 publications

References 47 publications

Synthetic Control of Protein Degradation during Cell Proliferation and Developmental Processes

Synthetic Control of Protein Degradation during Cell Proliferation and Developmental Processes

Degrons of yeast and mammalian ornithine decarboxylase enzymes make potent combination for regulated targeted protein degradation

Structural and Functional Characterisation of the Domains of Ubiquitin-Activating Enzyme (E1) of Saccharomyces cerevisiae

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