Dataset Information

SCFCyclin F targets the metabolic protein Sirtuin 5 for ubiquitination

ABSTRACT: Cell cycle transitions are regulated by a number of coordinating signaling events. The ubiquitin-proteasome system is an important part of this cell cycle signaling. Cyclin F, a substrate adapter for the SCF (SKP1/CUL1/F-box) E3 ubiquitin ligase, is an important regulator of cell cycle progression. However, identifying SCFCyclin F substrates important for the G1-S transition and other cell cycle events has remained technically challenging. Since Cyclin F overexpression rescues a yeast mutant in the cdc4 gene, we considered the possibility that other genes that genetically modify cdc4 mutant lethality could also encode Cyclin F substrates. We identified the mitochondrial and cytosolic deacylating enzyme Sirtuin 5 (SIRT5) as a novel Cyclin F substrate. SIRT5 has been implicated in a number of metabolic processes, but its connection to the cell cycle is not known. Here, we demonstrate the ability of Cyclin F to interact with, ubiquitinate and control the abundance and stability of SIRT5. Since previously identified Cyclin F targets are important for cell cycle progression, we hypothesized that SIRT5 would influence cell cycle progression. We show SIRT5 CRISPR knockout results in a diminished G1 population, and subsequent increase in both S and G2/M. Proteomic analyses reveal CDK signaling changes congruent with the cell cycle changes in SIRT5 knockout cells. Together these data demonstrate SIRT5 is regulated by Cyclin F and suggest a connection between SIRT5, cell cycle regulation, and metabolism.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Matthew D. Hirschey

PROVIDER: PXD019396 | JPOST Repository | Tue Nov 17 00:00:00 GMT 2020

REPOSITORIES: jPOST

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	2019-06-12_Sirt5CRISPRFeeding_WT-TMT11-InputF1_AMWT1-1.raw	Raw
	2019-06-12_Sirt5CRISPRFeeding_WT-TMT11-InputF1_AMWT1-2.raw	Raw
	2019-06-12_Sirt5CRISPRFeeding_WT-TMT11-InputF1_AMWT1-3.raw	Raw
	2019-06-12_Sirt5CRISPRFeeding_WT-TMT11-InputF1_AMWT1-4.raw	Raw
	2019-06-12_Sirt5CRISPRFeeding_WT-TMT11-InputF1_AMWT1-5.raw	Raw

Items per page:

1 - 5 of 20

Publications

Sirtuin 5 Is Regulated by the SCFCyclin F Ubiquitin Ligase and Is Involved in Cell Cycle Control.

Mills Christine A CA Wang Xianxi X Bhatt Dhaval P DP Grimsrud Paul A PA Matson Jacob Peter JP Lahiri Debojyoti D Burke Daniel J DJ Cook Jeanette Gowen JG Hirschey Matthew D MD Emanuele Michael J MJ

Molecular and cellular biology 20210125 2

The ubiquitin-proteasome system is essential for cell cycle progression. Cyclin F is a cell cycle-regulated substrate adapter F-box protein for the Skp1, CUL1, and F-box protein (SCF) family of E3 ubiquitin ligases. Despite its importance in cell cycle progression, identifying cyclin F-bound SCF complex (SCFCyclin F) substrates has remained challenging. Since cyclin F overexpression rescues a yeast mutant in the cdc4 gene, we considered the possibility that ...[more]

PMID: 33168699

Similar Datasets

Project description:In the budding yeast Saccharomyces cerevisiae, transcription factors (TFs) regulate the periodic expression of many genes during the cell cycle, including gene products required for progression through cell-cycle events. Experimental evidence coupled with quantitative models suggest that a network of interconnected TFs is capable of regulating periodic genes over the cell cycle. Importantly, these dynamical models were built on transcriptomics data and assumed that TF protein levels and activity are directly correlated with mRNA abundance. To ask whether TF transcripts match protein expression levels as cells progress through the cell cycle, we applied a multiplexed targeted mass spectrometry approach (parallel reaction monitoring) on synchronized populations of cells. We found that protein expression of many TFs and cell-cycle regulators closely followed their respective mRNA transcript dynamics in cycling wild-type cells. Discordant mRNA/protein expression dynamics were also observed for a subset of cell-cycle TFs and for proteins targeted for degradation by E3 ubiquitin ligase complexes such as SCF (Skp1/Cul1/F-box) and APC/C (anaphase-promoting complex/cyclosome). We further profiled mutant cells lacking B-type cyclin/CDK activity (clb1-6), where oscillations in ubiquitin ligase activity, cyclin/CDKs, and cell-cycle progression are halted. We found that a number of proteins were no longer periodically degraded in clb1-6 mutants compared to wild type, highlighting the importance of post-transcriptional regulation. Finally, the TF complexes responsible for activating G1/S transcription (SBF and MBF) were more constitutively expressed at the protein level than their periodic mRNA expression levels in both wild-type and mutant cells. This comprehensive investigation of cell-cycle regulators reveals that multiple layers of regulation (transcription, protein stability, and proteasome targeting) affect protein expression dynamics during the cell cycle.

Dataset Information

SCFCyclin F targets the metabolic protein Sirtuin 5 for ubiquitination

Dataset's files

Publications

Sirtuin 5 Is Regulated by the SCF<sup>Cyclin F</sup> Ubiquitin Ligase and Is Involved in Cell Cycle Control.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets