RT Journal Article
SR Electronic
T1 A CRISPR-based genome-wide screen for adipogenesis reveals new insights into mitotic expansion and lipogenesis
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.07.13.201038
DO 10.1101/2020.07.13.201038
A1 Keren I. Hilgendorf
A1 Carl T. Johnson
A1 Kyuho Han
A1 Atefeh Rabiee
A1 Janos Demeter
A1 Ran Cheng
A1 Yingdi Zhu
A1 Zewen Jiang
A1 Katrin J. Svensson
A1 Michael C. Bassik
A1 Peter K. Jackson
YR 2020
UL http://biorxiv.org/content/early/2020/07/14/2020.07.13.201038.abstract
AB In response to excess nutrients, white adipose tissue expands by both generating new adipocytes and by upregulating lipogenesis in existing adipocytes. Here, we performed a genome-wide functional genomics screen to identify regulators of adipogenesis in the mouse 3T3-L1 cell model. The pooled screening strategy utilized FACS to isolate populations based on lipid content by gating for fluorescence intensity of the lipophilic, green fluorescent BODIPY 493/503 dye. Additionally, this approach categorized if genes functioned during mitotic expansion or lipogenesis. Cellular mechanisms regulating the rates of protein translation and protein stability were found to be critical for adipogenesis and lipogenesis. These mechanisms were further supported by proteomic analyses, which demonstrated that many changes in protein abundance during 3T3-L1 adipogenesis were not driven by transcription. Within these themes, we illustrate that hypusination is critical for translating adipogenic inducers of mitotic expansion and that the neddylation/ubiquitin pathway modulates insulin sensitivity to regulate lipogenesis.Competing Interest StatementThe authors have declared no competing interest.