XB-ART-60505
Mol Biol Cell
2024 Mar 01;353:ar39. doi: 10.1091/mbc.E23-03-0084.
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Label-free proteomic comparison reveals ciliary and nonciliary phenotypes of IFT-A mutants.
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DIFFRAC is a powerful method for systematically comparing proteome content and organization between samples in a high-throughput manner. By subjecting control and experimental protein extracts to native chromatography and quantifying the contents of each fraction using mass spectrometry, it enables the quantitative detection of alterations to protein complexes and abundances. Here, we applied DIFFRAC to investigate the consequences of genetic loss of Ift122, a subunit of the intraflagellar transport-A (IFT-A) protein complex that plays a vital role in the formation and function of cilia and flagella, on the proteome of Tetrahymena thermophila. A single DIFFRAC experiment was sufficient to detect changes in protein behavior that mirrored known effects of IFT-A loss and revealed new biology. We uncovered several novel IFT-A-regulated proteins, which we validated through live imaging in Xenopus multiciliated cells, shedding new light on both the ciliary and non-ciliary functions of IFT-A. Our findings underscore the robustness of DIFFRAC for revealing proteomic changes in response to genetic or biochemical perturbation.
???displayArticle.pubmedLink??? 38170584
???displayArticle.pmcLink??? PMC10916875
???displayArticle.link??? Mol Biol Cell
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F31 DE029114 NIDCR NIH HHS, R01 HD085901 NICHD NIH HHS , R35 GM122480 NIGMS NIH HHS , R35 GM140813 NIGMS NIH HHS
Genes referenced: ak8 ccdc157 cfap45 ift122 pccb pip5kl1 pkm rfx2 sort1
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