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| Author(s): |
Deivid Lucas dos Santos Migueleti
Total Authors: 1
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| Document type: | Master's Dissertation |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia |
| Defense date: | 2012-04-06 |
| Examining board members: |
Jörg Kobarg;
Deborah Schechtman;
Claudio Chrysostomo Werneck
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| Advisor: | Jörg Kobarg |
| Abstract | |
The protein UNC-76 was identified as necessary for fasciculation and elongation of axons of the worm Caenorhabditis elegans during development of the nervous system. The mammalian homologue FEZ1 is mostly expressed in neuronal tissues and FEZ1 knockout mice present behavior abnormalities that resemble neurological disorders. The role of FEZ1 in the development of the nervous system seems to lie in its association with cytoskeletal elements and signaling pathways (e.g., PKC?, E4B, DISC1) regulating axon outgrowth and cell polarization. The studies of our group have shown that FEZ1 is a hub, able to interact with more than 50 partners through its coiled-coil domains. Furthermore, overexpression of FEZ1 in HEK293 cells causes the appearance of flower-like nuclei, a common phenotype to certain types of leukemia. In this work the role of FEZ1 in the mechanisms of flower-like nuclei formation and functional consequences of its overexpression on cell viability were investigated, attempting to extrapolate this model for leukemias. In silico analysis of several leukemias showed that FEZ1 is overexpressed in AML patients and that this may relate to the occurrence of 11q23/MLL genetic fusion. FEZ1 expression in leukemic THP-1 cells was detected by Western blotting, but the expression in PBMCs of leukemic patients still lacks empirical evidence. To assess the functional consequences of overexpression, cell lineage with stable and inducible expression of FEZ1 was obtained and used in proliferative assays. However, it was not observed any differences between lineages expressing FLAG-FEZ1 fusion protein or FLAG tag alone. IP-MS assay using these lineages identified proteins whose interaction with FEZ1 could be modulated by the activity of PKCs. Finally, cotransfection of C-terminal coiled-coils and FEZ1 full-length decreases flower-like nuclei formation to nearly 40%. Transfection with FEZ1nocys mutant containing five substituted cysteines did not play the same, but further experiments are needed to determine the potential synergism these two components may have on this phenomenon (AU) | |
| FAPESP's process: | 10/02322-7 - FEZ1 and the "flower-like" phenotype formation: mechanisms and consequences in cancer. |
| Grantee: | Deivid Lucas dos Santos Migueleti |
| Support Opportunities: | Scholarships in Brazil - Master |