The central domain of yeast transcription factor Rpn4 facilitates degradation of reporter protein in human cells

•We showed proteasomal hydrolysis of yeast transcription factor Rpn4 in human cells•N-terminal degron of the yRpn4 cannot serve as an effective degron in human cells•We have shown that central domain of the yRpn4 contains efficient interspecies degron•Discovered degron can be used to induce degradat...

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Published in:	FEBS letters Vol. 588; no. 20; pp. 3713 - 3719
Main Authors:	Morozov, A.V., Spasskaya, D.S., Karpov, D.S., Karpov, V.L.
Format:	Journal Article
Language:	English
Published:	England Elsevier B.V 16-10-2014
Subjects:	AFP Alpha-fetoprotein alpha-Fetoproteins - metabolism CMV cytomegalovirus DNA-Binding Proteins - chemistry DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism GFP green fluorescent protein HEK293 Cells Humans ODC ornithine decarboxylase Proteasome Proteasome Endopeptidase Complex - metabolism Protein degradation signals Protein Structure, Tertiary Proteolysis Rpn4 Saccharomyces cerevisiae Proteins - chemistry Saccharomyces cerevisiae Proteins - genetics Saccharomyces cerevisiae Proteins - metabolism Transcription Factors - chemistry Transcription Factors - genetics Transcription Factors - metabolism ubiquitin GFP Rpn4 Protein degradation signals AFP Ub CMV ODC Proteasome
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Summary:	•We showed proteasomal hydrolysis of yeast transcription factor Rpn4 in human cells•N-terminal degron of the yRpn4 cannot serve as an effective degron in human cells•We have shown that central domain of the yRpn4 contains efficient interspecies degron•Discovered degron can be used to induce degradation of target proteins in human cells Despite high interest in the cellular degradation machinery and protein degradation signals (degrons), few degrons with universal activity along species have been identified. It has been shown that fusion of a target protein with a degradation signal from mammalian ornithine decarboxylase (ODC) induces fast proteasomal degradation of the chimera in both mammalian and yeast cells. However, no degrons from yeast-encoded proteins capable to function in mammalian cells were identified so far. Here, we demonstrate that the yeast transcription factor Rpn4 undergoes fast proteasomal degradation and its central domain can destabilize green fluorescent protein and Alpha-fetoprotein in human HEK 293T cells. Furthermore, we confirm the activity of this degron in yeast. Thus, the Rpn4 central domain is an effective interspecies degradation signal.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0014-5793 1873-3468
DOI:	10.1016/j.febslet.2014.08.017