Determination of the structure of the DWNN-containing protein from the protozoon Encephalitozoon cuniculi

Determination of the structure of the DWNN-containing protein from the protozoon <em>Encephalitozoon cuniculi</em>

using NMR or X-ray crystallography

Encephalitozoon cuniculi has one of the smallest eukaryotic genomes characterised, and is a possible candidate for the so-called "minimal" eukaryotic genome. Many of the genes are highly compactified versions of their orthologues in other organisms () ()The DWNN-containing protein in this organism consists of the DWNN domain, the Zinc finger and the Ring finger, and is therefore similar to the form found in yeast, which is known to be involved in mRNA processing. However it is only 260 amino acids long, as compared to 440 in yeast, which makes it suitable for structure determination by either NMR or X-ray. The structures of the individual domains from human are currently being determined by NMR and comparisons with the complete protein from E. cuniculi will allow the structure of the human isoform to be modelled. It may also answer the question as to whether the additional residues serve any function in humans, or whether they are simply redundant.

At our request the gene has been cloned into the cloning vector pGEM-T-easy by the French group responsible for sequencing the genome. The project will involve amplification of the gene using PCR and cloning into a GST expression vector, optimisation of expression and solubility conditions, and preliminary NMR studies and crystallisation trials. Depending on the success of the former, NMR or X-ray data will be collected and the structure determined in collaboration with either David or Mohammed.

References

  1. Roger, A. J., & Silberman, J. D. (2002). Cell evolution: mitochondria in hiding. Nature, 418(6900), 827-9. PMID: 12192393 ()
  2. Katinka, M. D., Duprat, S., Cornillot, E., Méténier, G., Thomarat, F., Prensier, G., . . . Vivarès, C. P. (2001). Genome sequence and gene compaction of the eukaryote parasite Encephalitozoon cuniculi. Nature, 414(6862), 450-3. PMID: 11719806 ()

Bibliography