Instruct: Identify the main category of Biorxiv papers based on the titles and abstracts
Query: Visual detection of binary, ternary, and quaternary protein-protein interactions in fission yeast by Pil1 co-tethering assay Protein-protein interactions are vital for executing nearly all cellular processes. To facilitate the detection of protein-protein interactions in living cells of the fission yeast Schizosaccharomyces pombe, here we present an efficient and convenient method termed the Pil1 co-tethering assay. In its basic form, we tether a bait protein to mCherry-tagged Pil1, which forms cortical filamentary structures, and examine whether a GFP-tagged prey protein colocalizes with the bait. We demonstrate that this assay is capable of detecting pairwise protein-protein interactions of cytosolic proteins, transmembrane proteins, and nuclear proteins. Furthermore, we show that this assay can be used for detecting not only binary protein-protein interactions, but also ternary and quaternary protein-protein interactions. Using this assay, we systematically characterized the protein-protein interactions in the Atg1 complex and in the phosphatidylinositol 3-kinase (PtdIns3K) complexes and found that Atg38 is incorporated into the PtdIns3K complex I via an Atg38-Vps34 interaction. Our data show that this assay is a useful and versatile tool and should be added to the routine toolbox of fission yeast researchers.
26
The categories of this paper are: biochemistry
1
The categories of this paper are: bioinformatics
18
The categories of this paper are: systems
25
The categories of this paper are: bioengineering
19
The categories of this paper are: epidemiology
12
The categories of this paper are: behavior
27
The categories of this paper are: pathology
16
The categories of this paper are: microbiology
15
The categories of this paper are: developmental
4
The categories of this paper are: neuroscience
17
The categories of this paper are: molecular
14
The categories of this paper are: biophysics
29
The categories of this paper are: education
11
The categories of this paper are: cognition
3
The categories of this paper are: evolutionary
28
The categories of this paper are: communication