Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transfer of 5d old Col-0 and WEE1 KO seedlings to medium supplemented with 2mM HU for 0h-5h-24h

ABSTRACT: Col-0 and WEE1KO (wee1-1 and wee1-2) were germinated on control medium on a nylon mesh and transferred 5 days after germination to medium supplemented with 2 mM HU. Samples were harvested at three different time points after transfer: 0 h, 5 h, and 24 h. All sampling points were performed in four independent experiments for Col-0, two independent experiments for wee1-1 and 2 independent experiments for wee1-2. For each time point in each experiment ﾱ50 root tips were collected and frozen in liquid nitrogen. RNA was extracted from root tissue with TriZol reagent (Invitrogen) and purified with RNneasy kit (Qiagen). The RNA of two independent experiments for Col-0 and WEE1KO were subsequently pooled and used for
microarray analysis.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Toon Cools

PROVIDER: E-MEXP-3053 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The Arabidopsis thaliana checkpoint kinase WEE1 protects against premature vascular differentiation during replication stress.

Cools Toon T Iantcheva Anelia A Weimer Annika K AK Boens Shannah S Takahashi Naoki N Maes Sara S Van den Daele Hilde H Van Isterdael Gert G Schnittger Arp A De Veylder Lieven L

The Plant cell 20110415 4

A sessile lifestyle forces plants to respond promptly to factors that affect their genomic integrity. Therefore, plants have developed checkpoint mechanisms to arrest cell cycle progression upon the occurrence of DNA stress, allowing the DNA to be repaired before onset of division. Previously, the WEE1 kinase had been demonstrated to be essential for delaying progression through the cell cycle in the presence of replication-inhibitory drugs, such as hydroxyurea. To understand the severe growth a ...[more]

PMID: 21498679

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Project description:Background: Atherosclerosis leads high mortality, highlighting an urgent need for new therapeutic strategies. Protein kinases orchestrate multiple cellular events in atherosclerosis and may provide new therapeutic targets for atherosclerosis. Here, we identified a protein kinase, WEE1 G2 checkpoint kinase (WEE1), promoting inflammatory response in atherosclerosis. Methods: The ApoE-/- mice without macrophage-specific WEE1 deletion (ApoE-/-WEE1f/f) and ApoE-/- mice with macrophage-specific WEE1 deletion (ApoE-/-WEE1MCKO) were generated using bone marrow transplantation. These mice and WEE1 inhibitor MK1775 were used in a high-fat diet (HFD)-induced atherosclerotic model. Human atherosclerotic tissues, mouse primary peritoneal macrophages (MPMs), 293T cells, and recombinant proteins were utilized to investigate the pathogenic role and underlying mechanisms of WEE1. Results: We identified up-regulated p-WEE1 in macrophages in atherosclerotic lesions of HFD-fed ApoE-/- mice. Transcriptome sequencing analysis indicated that WEE1 promotes oxLDL-induced inflammation in macrophages. We then demonstrated that macrophage-specific deletion or pharmacological inhibition of WEE1 attenuates atherosclerosis by reducing inflammation both in vivo and in vitro. The overexpression of wild-type WEE1 or activating-mutant WEE1, but not inactivating-mutant WEE1, exacerbates inflammation in macrophages. Mechanistically, transcriptome sequencing analysis and co-immunoprecipitation followed by quantitative proteomics analysis identified p65 as a binding protein of WEE1. We confirmed that WEE1 directly interacts with the RHD domain of p65 via kinase domain and phosphorylates p65 at S536, thereby facilitating subsequent NF-κB activation and inflammatory response in macrophages. Conclusions: Our findings demonstrated that macrophage WEE1 promotes inflammatory atherosclerosis by directly binding to p65 and phosphorylate it at S536. This study provides WEE1 as a new p65 regulator in inflammation and a potential therapeutic target for atherosclerosis.

Dataset Information

Transfer of 5d old Col-0 and WEE1 KO seedlings to medium supplemented with 2mM HU for 0h-5h-24h

Publications

The Arabidopsis thaliana checkpoint kinase WEE1 protects against premature vascular differentiation during replication stress.

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