AI-facilitated multi-omics reveal TRF-controlled propionylation in determining protein homeostasis

ABSTRACT: Firstly, we systematically identified rhythmically modified Kpr sites, through time-series propionylomic profiling of fly heads. Wild-type (WT) and per0 flies were entrained under light/dark cycles (LD) and then released into constant darkness (DD), respectively. On the first day of DD, whole heads of WT flies were collected every 6 hours, including Zeitgeber time 0 (ZT0), ZT6, ZT12 and ZT18, whereas per0 fly heads were collected at ZT6 and ZT18, respectively. For each sample, whole-cell lysates were digested by trypsin and labeled with one of the tandem mass tag (TMT) 6-plex reagents. Then, propionylated peptides were isolated by immuno-affinity enrichment using the pan-propionylation antibody. The 6 samples were mixed with equal amounts and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Secondly, metabolomic profiling was conducted using TRF-WT and TRF-H2BK17A fly heads collected at ZT9 and ZT21. After the preprocessing including homogenization and different reagents mixture followed by centrifugation. All samples were analyzed by the LC-MS/MS system. Additionally, Immunoprecipitaion and LC-MS/MS analysis was perform with fly heads using H2BK17pr antibody to evaluate the its enrichment capacity.

ORGANISM(S): Drosophila Melanogaster

SUBMITTER: Yu Xue  

PROVIDER: PXD040987 | iProX | Mon Mar 20 00:00:00 GMT 2023

REPOSITORIES: iProX