Dataset Information

Inducible synthetic protein degradation system for phenotype control

ABSTRACT: Synthetic control of cell factory phenotypes is needed for redirecting resources from cell growth to production. To achieve that ClpXP proteasome was introduced to yeast Saccharomyces cerevisiae under control of TetON induction. Growth essential (in synthetic defined medium) metabolic enzymes Acc1, Aro1, Hom3 were targeted to the ClpXP proteasome by introducing short ssrA tags. Inducing the ClpXP proteasome with doxycycline during exponential growth phase ceased the growth.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Frank Stein

LAB HEAD: Paula Jouhten

PROVIDER: PXD034451 | Pride | 2023-05-10

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	Ernie_191105_P1125_MR_PJ_TP_TMT11_rep1_fr1.raw	Raw
	Ernie_191105_P1125_MR_PJ_TP_TMT11_rep1_fr10.raw	Raw
	Ernie_191105_P1125_MR_PJ_TP_TMT11_rep1_fr11.raw	Raw
	Ernie_191105_P1125_MR_PJ_TP_TMT11_rep1_fr12.raw	Raw
	Ernie_191105_P1125_MR_PJ_TP_TMT11_rep1_fr2.raw	Raw

Items per page:

1 - 5 of 82

Publications

Inducible Synthetic Growth Regulation Using the ClpXP Proteasome Enhances cis,cis-Muconic Acid and Glycolic Acid Yields in <i>Saccharomyces cerevisiae</i>.

Kakko Natalia N Rantasalo Anssi A Koponen Tino T Vidgren Virve V Kannisto Matti M Maiorova Natalia N Nygren Heli H Mojzita Dominik D Penttilä Merja M Jouhten Paula P

ACS synthetic biology 20230328 4

Engineered microbial cells can produce sustainable chemistry, but the production competes for resources with growth. Inducible synthetic control over the resource use would enable fast accumulation of sufficient biomass and then divert the resources to production. We developed inducible synthetic resource-use control over<i>Saccharomyces cerevisiae</i> by expressing a bacterial ClpXP proteasome from an inducible promoter. By individually targeting growth-essential metabolic enzymes Aro1, Hom3, a ...[more]

PMID: 36976676

Similar Datasets

Project description:Intracellular bacterial pathogens inject a cocktail of effector proteins into host cells to hijack diverse cellular processes and promote their survival and proliferation. Salmonella enterica serovar Typhimurium (STm) uses two Type 3 secretion system (T3SS) to deliver a suite of effector proteins into host cells, however, knowledge of their host-targets remains sparse. To systematically map STm effector-host protein-protein interactions (PPIs) during active infection, we generated a library of 32 STm strains expressing affinity-tagged effector proteins from their endogenous chromosomal loci. Tagged-effector strains were then used to infect macrophages or epithelial cells, and PPIs were detected by Affinity-Purification Quantitative Mass-Spectrometry (AP-QMS) under both native and cross-linking conditions. We recovered 25 previously described effector-host PPIs, along with 421 novel interactions across the two cell lines tested, as well as several effector-effector PPIs. While effectors converged on specific host cellular processes, which varied across cell types, most had multiple host targets. Using reciprocal co-immunoprecipitations, we could validated 13 out of 22 new PPIs, demonstrating an accuracy of at least 59% for the AP-QMS approach. To illustrate the utility of this resource for discovering novel infection biology, we first showed that SseJ and SseL cooperate to control cholesterol transport at the Salmonella Containing Vacuole (SCV) via the Niemann-Pick C1 protein (NPC1). Second, we uncovered that PipB directly binds and recruits the organelle contact site protein PDZD8 to the SCV. Third, we elucidated a novel mechanism whereby the effector kinase SteC promotes host actin bundling, by directly interacting and phosphorylating formin-like proteins. Overall, we provide a systems-wide host-bacterial pathogen physical interactome resource, to our knowledge the first in an infection context, with broad implications for effector function and cooperation.