Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: Quantitative phosphoproteomics reveals regulatory state dependent efficacy of chemical Cdk1 inhibition and distinct Cdk5 complexes as novel Wee1 substrates Wee1 kinase plays a central role in the eukaryotic cell cycle via its well-known negative regulation of Cdk1 activity at the G2/M transition, preventing progression into mitosis until DNA replication and/or DNA damage repair is complete. Recent genetic evidence in yeast, flies and human cells have suggested additional functions of Wee1 in mitosis and during mitotic exit, respectively. To discover new candidate substrates of Wee1 kinase, we used SILAC-based phosphoproteomics and selective chemical inhibition to quantitatively compare phosphorylation site abundances in the presence and absence of Wee1 activity. Unexpectedly, we uncovered a role for the Wee1-dependent phosphorylation of Cdk1-cyclin B at tyrosine 15 (Y15) in facilitating chemical inhibition of Cdk1-cyclin B by the inhibitor RO3306. Thermal shift stability assays demonstrated greater binding affinity of RO3306 for Y15-phosphorylated Cdk1-cyclin B versus unphosphorylated complex, providing an additional molecular basis for the observed Wee1 inhibitor-based toxicity in human cells. In addition, our experiments identified Cdk5-CABLES and Cdk5-cyclin B as novel substrates of Wee1 during chemically induced exit from mitosis. Collectively, these experiments facilitate a greater understanding of the Wee1-Cdk1 signaling axis and uncover new candidate substrates for Wee1.
22
The categories of this paper are: physiology
25
The categories of this paper are: bioengineering
4
The categories of this paper are: neuroscience
16
The categories of this paper are: microbiology
24
The categories of this paper are: cancer
6
The categories of this paper are: plant
28
The categories of this paper are: communication
19
The categories of this paper are: epidemiology
12
The categories of this paper are: behavior
31
The categories of this paper are: paleontology
10
The categories of this paper are: and
8
The categories of this paper are: genomics
1
The categories of this paper are: bioinformatics
30
The categories of this paper are: scientific
0
The categories of this paper are: genetics
29
The categories of this paper are: education