Introduction
Methylation is a common post-translational modification of proteins, which affects their function, and thus may impact various intracellular processes. Methyltransferases (MTases) are the enzymes that catalyse methylation reactions, and >200 such enzymes are present in humans. About 1/4 of these are so called SET proteins that are largely known for methylating lysine residues in the flexible tails of histone proteins, contributing to epigenetic regulation. However, SET enzymes can also methylate lysines in non-histone proteins. Recently, we became interested in one of the, so far, largely uncharacterized SET proteins (SETX) that is involved in cell quiescence and organ regeneration. Although SETX was reported to mediate methylation of specific lysine residues in histones and some other proteins, the underlying biochemical data are weak and indirect, thus requiring additional verification.
Aims
The aim of this Master project is to better characterize the MTase activity of SETX, both in vitro and in vivo. The particular goal is to identify novel protein substrate(s) of SETX, and establish a direct link between methylation of these substrate(s) and the biological role of SETX.
Methods
The potential student will learn how to apply fundamental molecular and cell biology techniques to make fundamental scientific discoveries. In particular, the student will learn molecular cloning and mutagenesis, and how to genetically manipulate human cells to generate knock-out (KO) cell lines and complementing them with a gene of interest. A great emphasis will be put on establishing functional consequences of gene KO and complementation. The student will also learn how to express and purify recombinant proteins from Escherichia coli, and how to perform methylation reactions in vitro. Methylation will also be studied in vivo, using mass spectrometry to verify the presence of methyl groups in proteins isolated from cellular material. Of note, similar strategies were previously used in Falnes group for identification of substrates of several other human MTases (PMID: 36997089).
Working environment
We are a tight-knit team of scientists who enjoy working hard, while having a lot of fun! We publish frequently in highly merited journals, such as Nature Communications and Nucleic Acids Research.
Put your talent into action and join our group!
Contact us: J?drzej Ma?ecki (j.m.malecki@ibv.uio.no) or P?l Falnes (pal.falnes@ibv.uio.no).