Project description:Microproteins are generated from small open reading frames (sORFs) and turn out to play various vital biological functions. As an essential biological event of eukaryotic cells, the cell cycle is involved in cell replication and division. For such a highly regulated event, microproteins associated with cell cycle regulation remained unclarified. Utilizing a combination of bottom-up and top-down proteomics, we analyzed microproteins at specific cell cycle stages of Hep3B cells. A total of 657 microproteins were identified under three cell cycle stages, including 151 in the G0/G1 stage, 163 in the S stage, and 132 in the G2/M stage. The annotation of these microproteins showed their cell cycle-specific functions, such as translation, nuclear assembly, chromatin organization, G2/M transition of the mitotic cell cycle. Meanwhile, more than 50 % of identified microproteins were ncRNA encoded. These non-annotated novel microproteins contain several function domains, such as nucleoside diphosphate kinase (NDK) domain, high mobility group (HMG) domain, and DNA-binding domain. That suggested the potential functions of these novel microproteins in specific cell cycle stages. This study presented a large-scale profile of microproteins at different cell cycle stages from Hep3B and may provide new perspectives on the regulation mechanism of the cell cycle.
Project description:In this study, we aimed to study the gene expression patterns at single cell level across the different cell cycle stages in mESC. We performed single cell RNA-Seq experiment on mESC that were stained with Hoechst 33342 and Flow cytometry sorted for G1, S and G2M stages of cell cycle. Single cell RNA-Seq was performed using Fluidigm C1 system and libraries were generated using Nextera XT (Illumina) kit.
Project description:This study is aimed to decipher the gene expression patterns that exist across the different cell cycle stages at level of individual cells and their inherent heterogeneity. Conventional studies in yeast and human cells have shown coordination between gene expression and cell-cycle. However such a study for mouse ES cells (mES) and their differentiation has not been performed at a single cell level. In this study, we obtain pure mES cells from different cell cycle stages using FACS and apply single cell RNA-sequencing using the Fluidigm C1 system. This work will decipher and unravel the transcriptional patterns that coordinated with cell cycle progression at a single cell level.
Project description:au13-11_gravity - gravity - Cell cycle and cell proliferation are decoupled under altered gravity conditions. We have previously shown that semisolid cell cultures of Arabidopsis suffer overall genome changes in response to altered gravity and also that cell cycle stages duration is altered. By using synchronized cell cultures we will demonstrate the precise alterations in cell cycle duration and also the transcriptional signature in any of them. - Experiments consists on exposures of Arabidopsis cell cultures to 1g control/simulated microgravity (RPM) conditions. Asynchronous cells exposed for 14 h + Syncronous populations choosen to have an enrichment of cell cycle phases were used (being T7/T10 samples on G2 phase, T14/T16 samples on G1 phase).
Project description:au13-11_gravity - gravity - Cell cycle and cell proliferation are decoupled under altered gravity conditions. We have previously shown that semisolid cell cultures of Arabidopsis suffer overall genome changes in response to altered gravity and also that cell cycle stages duration is altered. By using synchronized cell cultures we will demonstrate the precise alterations in cell cycle duration and also the transcriptional signature in any of them. - Experiments consists on exposures of Arabidopsis cell cultures to 1g control/simulated microgravity (RPM) conditions. Asynchronous cells exposed for 14 h + Syncronous populations choosen to have an enrichment of cell cycle phases were used (being T7/T10 samples on G2 phase, T14/T16 samples on G1 phase). 6 dye-swap - time course,treated vs untreated comparison
Project description:The host cell cycle is a major target for manipulation by viral pathogens, which often replicate in certain cell cycle stages. The prevalent pathogen human cytomegalovirus (HCMV), for instance, only replicates during G1 and therefore employs diverse mechanisms for regulating cell cycle progression. Here, we demonstrate that the human enzyme sirtuin 2 (SIRT2) is required for HCMV-mediated cell cycle dysregulation. By combining virology assays with mass pectrometry-based proteome, acetylome, and interactome analyses, we have defined a pro-virus function for SIRT2 in regulating the G1 to S phase transition.