The product of the c src proto oncogene, tyrosine kinase Src, is an essential regulator of cellular physiological processes ranging from cell adhesion, migration to mitogenic and anti apoptotic signaling. Thus, finding the precise regulation of Src in living cells is key to both understanding Src-dependent signal transduction and developing effective Src-targeted therapeutic approaches. Although Src is ubiquitously expressed, targeted disruption of c src in mice leads to only one major phenotype, osteopetrosis. This is the result of excessive accumulation of bone matrix caused by defective osteoclast function. The bones of mammals and birds differ in their inner structure and the role of Src in bird bone formation has not yet been analyzed.
The main research objective of this project is the elucidation of the role of Src in osteoclasts and the comparison of the specific role of Src in bone formation and osteoclast physiology between mammals and birds. The research will focus on description of the role of Src in bone formation in vivo and on osteoclasts physiology in vitro.
The in vivo experiments will include comparison of src / phenotype between murine and chicken. In collaboration with group of Dr. Hejnar (IMG, Prague) src / chicken will be prepared and analyzed for potential abnormalities in bone formation and bone resorbing activity of src / osteoclasts. These results will be compared to published phenotype of src / mice.
The in vitro experiments will focus on analyses of Src activation dynamics in correlation to bone resorbing activity of isolated osteoclasts. Src FRET biosensor technology developed in our lab will be used. Both chicken and mice osteoclasts expressing Src FRET biosensor will be prepared. The main focus will be on specific osteoclasts structures called sealing zones. These structures are responsible for bone resorption and cannot form when Src activity is inhibited. The dynamics of Src activation during formation of the sealing zones will be evaluated and correlated to bone resorption activity of the osteoclasts.
The candidate should have a strong background in molecular cloning, biochemical analyses, cell-based assays and experience with live-cell fluorescence microscopy. Experience with FRET imaging is of further advantage.
Koudelková L, Pataki AC, Tolde O, Pavlik V, Nobis M, Gemperle J, Anderson K, Brábek J, Rosel D. Novel FRET Based Src Biosensor Reveals Mechanisms of Src Activation and Its Dynamics in Focal Adhesions. Cell Chem Biol. 2019 Feb 21;26(2):255 268.e4. doi: 10.1016/j.chembiol.2018.10.024.
I declare that:
● co founding 1000 EUR/month is ensured
● project is approved by head of corresponding department
Co founding resources:
National Institute for Cancer Research (LX22NPO5102) – program EXCELES of the Ministry of Education, Youth, and Sports of the Czech Republic
Supervisor: Doc. RNDr. Daniel Rösel, Ph.D.
Department: Department of Cell Biology
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