Niels Erik Møllegaard
RNA and Gene Medicine Program
Panum, Bygning: 18.2.28
2200 København N
Genome structure and organisation, Protein-DNA interaction, Protein and DNA photooxidation, Protein purification method development.
DNA structural codes in genomes.
The sequence of genomic DNA contains information on several levels, ranging from simple coding of protein and RNA gene products over controlling the temporal and spatial expression of these to the physical structure and compaction (in nucleosomes and chromatin) of the genome in the cell nucleus, and therefore the evolutionary constraint on the genome sequence is multidimensional. Almost all genomic bioinformatics analyses focus on primary DNA sequence without taking into account or being able to interpret second and third order etc. effects of the DNA sequence, as for instance DNA helix conformation, electrostatics and flexibility which is as well “encoded” by the DNA sequence, and has direct impact on protein recognition and physical DNA properties. This project is aimed at understanding these sequence derived effects on DNA helix conformation, in particular in terms of genome function and thus evolutionary impact, as well as developing bioinformatics tools which allow direct sequence based genomic interrogation of DNA helix conformation and minor groove electrostatics, thereby obtaining a deeper genome wide understanding of the functional importance (also in terms of disease) of these.
Novel Biotechnology-based method for synthesis of amidated peptide pharmaceuticals.
In this project we propose an entirely new strategy for producing amidated peptides from E. coli fermentation both solving enrichment and amidation efficiency of the peptide. The technology is based on uranyl (UO2 2+) photo-cleavage of a novel C-terminal peptide tag that specifically binds to uranyl with high affinity. UV irradiation efficiently induces a site-specific cleavage of the peptide tag yielding a C-terminally amidated peptide and a uranyl-bound peptide tag. The purpose of this project is to develop a general method for production in E.coli of therapeutically relevant amidated peptides by developing a one-step peptide enrichment and amidation technology. This project is in collaboration with Proffs. Frank Kjeldsen and Thomas Jørgensen, University of Southern Denmark.
Photooxidation of DNA binding proteins.
It is widely established that exposure of cells to oxidants can result in cell dysfunction and death. Oxidation can result from many biological processes (e.g. inflammation, mitochondrial electron leakage, metabolic pathways, enzyme activity), as well as exposure to UV and high energy radiation, minerals, pesticides, and pollutants. We have shown that some DNA binding proteins are highly susceptible to biologically-relevant wavelengths and doses of UV light, with this resulting in protein aggregation and selective damage to the DNA to which the protein is bound. These exciting observations provide a new paradigm for modulating gene expression and growth of cells. This project is in collaboration with Prof. Michael Davies, BMI, Faculty of Health Sciences, KU.