TY - JOUR T1 - Proteomic analysis of cell cycle progression in asynchronous cultures, including mitotic subphases, using PRIMMUS JF - bioRxiv DO - 10.1101/125831 SP - 125831 AU - Tony Ly AU - Arlene Whigham AU - Rosemary Clarke AU - Alejandro Brenes-Murillo AU - Brett Estes AU - Patricia Wadsworth AU - Angus I. Lamond Y1 - 2017/01/01 UR - http://biorxiv.org/content/early/2017/04/09/125831.abstract N2 - The temporal regulation of protein abundance and post-translational modifications is a key feature of cell division. Recently, we analysed gene expression and protein abundance changes during interphase under minimally perturbed conditions (Ly et al. 2014; Ly et al. 2015). Here we show that by using specific intracellular immunolabeling protocols, FACS separation of interphase and mitotic cells, including mitotic subphases, can be combined with proteomic analysis by mass spectrometry. Using this PRIMMUS (PRoteomic analysis of Intracellular iMMUnolabeled cell Subsets) approach, we now compare protein abundance and phosphorylation changes in interphase and mitotic fractions from asynchronously growing human cells. We identify a set of 115 phosphorylation sites increased during G2, which we term ‘early risers’. This set includes phosphorylation of S738 on TPX2, which we show is important for TPX2 function and mitotic progression. Further, we use PRIMMUS to provide a proteome-wide analysis of protein abundance remodeling between prophase, prometaphase and anaphase. ER -