18.1. 2018, Bioenergetika

PhD project1:

Characterisation of TMEM70 protein function in mammalian mitochondria.

Our team was involved in the description and subsequent characterisation of TMEM70 protein acting as assembly factor for ATP synthase. Using mouse model of inducible TMEM70 knockout and cellular knockout models we want to elucidate true TMEM70 function and to search for potential new factors involved in regulation of biogenesis of ATP synthase and potentially other OXPHOS complexes.

Second part of the project will explore possible pharmacological complementation of ATP synthase assembly defects. Here TMEM70 represents promising target for establishing of new therapy as it is not absolutely essential for ATP synthase biogenesis and sufficient recovery of ATP production can be expected even after partial increase of ATP synthase content. The aim will be to screen for pharmacological activators of ATP synthesis on a model of TMEM70 knockout cells and subsequently test candidate compounds on a collection of patients’ fibroblasts cell lines harbouring mutations in TMEM70.

Candidate’s profile (requirements): MSc or MD degree in biochemistry, pharmacology or similar field. Candidates should have a good record of accomplishment in biochemistry, cell biology and molecular biology techniques.

Relevant publications:

1.   Cizkova A, Stranecky V, Mayr JA, Tesarova M, Havlickova V, Paul J, Ivanek R, Kuss AW, Hansikova H, Kaplanova V, Vrbacky M, Hartmannova H, Noskova L, Honzik T, Drahota Z, Magner M, Hejzlarova K, Sperl W, Zeman J, Houstek J, Kmoch S. TMEM70 mutations cause isolated ATP synthase deficiency and neonatal mitochondrial encephalocardiomyopathy. Nat Genet. 2008;40(11):1288-90.

2.   Kratochvilova H, Hejzlarova K, Vrbacky M, Mracek T, Karbanova V, Tesarova M, Gombitova A, Cmarko D, Wittig I, Zeman J, Houstek J. Mitochondrial membrane assembly of TMEM70 protein. Mitochondrion. 2014;15:1-9.

3.   Vrbacky M, Kovalcikova J, Chawengsaksophak K, Beck IM, Mracek T, Nuskova H, Sedmera D, Papousek F, Kolar F, Sobol M, Hozak P, Sedlacek R, Houstek J. Knockout of Tmem70 alters biogenesis of ATP synthase and leads to embryonal lethality in mice. Hum Mol Genet. 2016;25(21):4674-85.

Website: http://www.fgu.cas.cz/en/departments/bioenergetics

Supervisor: RNDr. Tomáš Mráček, Ph.D.

 

PhD project2:

Biogenesis of mammalian ATP synthase

Mitochondrial FoF1 ATP synthase is an enzyme, which shows good level of conservation already from bacteria. Yeast enzyme is very similar to the mammalian one, which led to assumption that the biogenesis should be conserved as well. However, identification of mammalian specific assembly factors (TMEM70) or accessory subunits (MLQ, Usmg5) as well as observation of intermediates not compatible with yeast assembly scheme (He J et al, 2017, PMID: 28289229) suggests rather different assembly pathway for the mammalian enzyme.

The aim of this project is to characterise assembly pathway for mammalian ATP synthase and explore the role of higher eucaryotes specific factors. For this, we have at hand cell knockout models for TMEM70, MLQ and Usmg5 subunits and animal knockouts for TMEM70 and Usmg5 and others should be produced by the successful candidate in the course of their studies.

 

Candidate’s profile (requirements): MSc or MD degree in (animal) physiology or similar. Candidates should have a good record of accomplishment in physiology and biochemistry. Willingness to work with laboratory animals is requirement, previous experience strong asset.

 

Supervisor: RNDr. Tomáš Mráček, Ph.D.

 

PhD project3:

omics approaches to deciphering adaptations of mitochondrial metabolism

Mitochondrial metabolism displays considerable plasticity and can adapt to external constrains, e.g. as we have demonstrated for leukemic cells treated with L-asparaginase. This project should combine data from measurements of cellular respiration/glycolysis with metabolomics data (generated at the IPHYS core) to pinpoint pro-survival pathways in various cancer cell lines that depend on functional mitochondria. Candidate should work closely with the metabolomics core staff and contribute to the development of fluxomic analysis of mitochondrial metabolism.

 

Candidate’s profile (requirements): MSc or equivalent degree in cell biology, biochemistry or analytical chemistry. Apart from the experimental “wet” work, candidates should be willing to learn data processing and analysis techniques and adapt them to project’s needs.

 

Relevant publications:

 1.        Hermanova I, Arruabarrena-Aristorena A, Valis K, Nuskova H, Alberich-Jorda M, Fiser K, Fernandez-Ruiz S, Kavan D, Pecinova A, Niso-Santano M, Zaliova M, Novak P, Houstek J, Mracek T, Kroemer G, Carracedo A, Trka J, Starkova J. Pharmacological inhibition of fatty-acid oxidation synergistically enhances the effect of l-asparaginase in childhood ALL cells. Leukemia. 2016;30(1):209-18.

2.         Pecinova A, Drahota Z, Kovalcikova J, Kovarova N, Pecina P, Alan L, Zima M, Houstek J, Mracek T. Pleiotropic Effects of Biguanides on Mitochondrial Reactive Oxygen Species Production. Oxid Med Cell Longev. 2017;2017:7038603

 

Supervisor:        RNDr. Alena Pecinová, Ph.D.