Core circadian clock transcription factor BMAL1 regulates mammary epithelial cell growth, differentiation, and milk component synthesis
Autor: | Jyothi Thimmapuram, Avi Shamay, Aridany Suarez-Trujillo, Shelby Cummings, Clare Aduwari, Phillip San Miguel, Ketaki Bhide, Sameer J. Mabjeesh, Katelyn Huff, Kelsey Teeple, Theresa Casey, Jennifer Crodian, Karen Plaut |
---|---|
Rok vydání: | 2021 |
Předmět: |
Physiology
Cellular differentiation Cell Circadian clock Biochemistry Ion Channels Mice Cell Signaling CRISPR-Associated Protein 9 Lactation Membrane Receptor Signaling Lipoprotein Receptors Gene Editing Multidisciplinary Physics ARNTL Transcription Factors Cell Differentiation Milk Proteins Hormone Receptor Signaling Cell biology Electrophysiology Circadian Rhythms medicine.anatomical_structure Gene Knockdown Techniques Physical Sciences Medicine Female Research Article Signal Transduction Transmembrane Receptors SOD3 Science Lipoproteins Biophysics Neurophysiology Anemia Sickle Cell Biology Mammary Glands Animal Cell surface receptor medicine Animals Transcription factor Gene Prolactin receptor Biology and Life Sciences Proteins Lipid metabolism Epithelial Cells Cell Biology Ligand-Gated Ion Channels Gene Expression Regulation CRISPR-Cas Systems Nuclear Receptor Signaling Chronobiology Neuroscience Developmental Biology |
Zdroj: | PLoS ONE PLoS ONE, Vol 16, Iss 8, p e0248199 (2021) |
ISSN: | 1932-6203 |
Popis: | The role the mammary epithelial circadian clock plays in gland development and lactation is unknown. We hypothesized that mammary epithelial clocks function to regulate mammogenesis and lactogenesis, and propose the core clock transcription factor BMAL1:CLOCK regulates genes that control mammary epithelial development and milk synthesis. Our objective was to identify transcriptional targets of BMAL1 in undifferentiated (UNDIFF) and lactogen differentiated (DIFF) mammary epithelial cells (HC11) using ChIP-seq. Ensembl gene IDs with the nearest transcriptional start site to peaks were explored as potential targets, and represented 846 protein coding genes common to UNDIFF and DIFF cells and 2773 unique to DIFF samples. Genes with overlapping peaks between samples (1343) enriched cell-cell adhesion, membrane transporters and lipid metabolism categories. To functionally verify targets, an HC11 line with Bmal1 gene knocked out (BMAL1-KO) using CRISPR-CAS was created. BMAL1-KO cultures had lower cell densities over an eight-day growth curve, which was associated with increased (pSod3). Q-PCR analysis also found lower expression of the putative targets, prolactin receptor (Prlr), Ppara, and beta-casein (Csn2). Findings support our hypothesis and highlight potential importance of clock in mammary development and substrate transport. |
Databáze: | OpenAIRE |
Externí odkaz: |