Příprava plazmidu, exprese a pilotní izolace proteinu MafK - interakčního partnera hemového senzoru Bach1

Mihalčin, Peter

Bakalárska práca Abstract Heme-sensing proteins are heme proteins to which heme serves as a signalling molecule. Association or dissociation of heme moiety and heme-sensing proteins influences various physiological functions, such as enzyme activity or gene expression regulated by these heme-sensing proteins. The main object of this thesis is heme-sensing protein Bach1 and its interaction partner, transcription factor MafK. Bach1 refers to the group of transcription factors involved in repression of gene expression. The target genes of Bach1 regulation are hemeoxygenase genes. Hemeoxygenase controls the excess free heme degradation. Due to the excess of free heme in the cell, Bach1-heme interaction inactivates Bach1 controlled repression of hemeoxygenase resulting in the free heme degradation. In the state of physiological free heme concentration, Bach1-heme interaction does not occur and activated Bach1 represses the hemeoxygenase expression via binding to the target gene enhancers. Bach1 is incapable of making efficient Bach1-DNA bonding by itself, therefore the transcription factor MafK is essential. Protein MafK modulates the Bach1-DNA binding by making the heterodimer formation Bach1-MafK, which binds to DNA. The first aim of this thesis is to summarize the recent knowledge about transcriptional...

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