The Mediator Complex has an essential function in regulating the transcription of several genes, working as a scaffold between transcription factors and polymerase II. It is composed of 26 subunits, with the kinase, or CDK subunit, being one of the most important for the mediator complex activation. In fact, when the kinase subunit is bound to the complex, binding between the mediator complex and polymerase II is hindered and therefore transcription is repressed. This suggests that the kinase complex and its components are essential for the expression of polymerase II transcripts. MED13 is part of the kinase subunit and high-throughput transcriptomic analysis showed MED13 as being upregulated in human skeletal muscle biopsies following an oral glucose tolerance test. The aim of this project is to understand the function of MED13 in human skeletal muscle metabolism and the relationship between MED13 knockdown and AMPK (AMP-activated protein kinase) function. Primary human skeletal muscles cells obtained from three healthy donors were cultured and transfection with MED13 siRNA was performed on differentiated myotubes. Metabolic assays such as fatty acid and glucose oxidation were performed as well as mitochondrial function. Quantitative PCR and western blot techniques were used to assess the pathways altered by MED13. Glucose uptake and fatty acid transporters were significantly increased upon MED13 silencing. Reduction in fatty acid oxidation and unchanged glucose oxidation suggest that MED13 favors substrate storage. Western blot analysis hinted on an increase in AMPK phosphorylation and in mitochondrial biogenesis. The production of ATP by mitochondria was also increased. In conclusion, MED13 plays an important role in human skeletal muscle substrate utilization for energy production.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:his-15527 |
| Date | January 2018 |
| Creators | Tiainen Losno, Emily Angela |
| Publisher | Högskolan i Skövde, Institutionen för biovetenskap, Karolinska Institutet |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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