The Svedberg Award 2007

Nico Dantuma
Department of Cell and Molecular Biology (CBM), Karolinska Institutet

Background
In 1992, I graduated in Biology and Medical Biology at the Free University in Amsterdam. As part of my undergraduate training, I had spent already more than one year at the bench being involved in two research projects dealing with enteropathogenic bacteria and estrogen-dependent growth of breast tumors at the Free University and the Netherlands Cancer Institute, respectively. It was during this time at the bench that I became really fascinated with science and in particular with doing research. Directly after my graduation, I started my PhD project at the Utrecht University where I studied how the African migratory locust (Locusta migratoria) mobilizes and transports lipids during their long distance flights resulting in the identification of a novel endocytotic lipoprotein receptor. In 1997, I defended my thesis and moved to Stockholm to join the laboratory of Maria Masucci, who was then at Microbiology and Tumor Biology center (MTC) of the Karolinska Institute. It was in the Masucci lab that I started to work on the ubiquitin/proteasome system, which is still the major focus of our research. As a postdoctoral fellow I studied how the Epstein Barr virus, which causes infectious mononucleosis and is linked to several forms of cancer, manipulates the ubiquitin/proteasome system. After my postdoctoral period, I established my own research group with a major focus on the ubiquitin/proteasome system in neurodegenerative disorders such as Alzheimer’s, Parkinson’s and Huntington’s disease. In 2003, I spent a sabbatical year in the group of Jacques Neefjes at the Netherlands Cancer Institute during which I became acquainted with various live cell imaging techniques. Shortly after my return to Stockholm, I moved my group to the department of Cell and Molecular Biology (CMB) also at the Karolinska Institute, where we continued our work on the ubiquitin/proteasome system in neurodegeneration as well as the development of new tools for following the ubiquitin/proteasome system in living cells. More recently, we started to study the role of the ubiquitin/proteasome system in DNA repair, a process that is highly relevant for our understanding of cancer.

Research
The ubiquitin/proteasome system is involved in many cellular processes. It is for example important for the cell cycle, programmed cell death, transcription, DNA repair, intracellular transport and, last but not least, for the destruction of misfolded or otherwise abnormal proteins. Best known is the role of the ubiquitin/proteasome system in regulated degradation of proteins but it also has less well understood non-proteolytic functions. Degradation is accomplished in a two step process. First, proteins that are destined for degradation are poly-ubiquitylated, which means that a long chain of a small protein called ubiquitin is being attached to the protein that will be degraded. Second, proteins with poly-ubiquitin chains bind to the proteasome, which is a large proteolytic complex that subsequently cuts the poly-ubiquitylated protein in small fragments. Regulated degradation of proteins is the perfect means to irreversibly inactivate regulatory proteins or to destroy abnormal and potentially dangerous proteins. It has become increasingly clear that there are different forms of ubiquitylation and many of those do not target proteins for degradation. For example, in transcription and DNA repair mono-ubiquitylation (attachment of a single ubiquitin) plays an important regulatory role without targeting proteins for degradation.

We have developed a number of tools that allows to analyse the functionality of the ubiquitin/proteasome system and to follow the dynamics of certain components of the system. Many neurodegenerative diseases are characterized by the presence of deposits of aggregated misfolded proteins in the affected brain regions. Since the ubiquitin/proteasome system is the primary pathway responsible for clearance of misfolded proteins, it has been proposed that problems with the ubiquitin/proteasome system may be a common nominator in neurodegenerative disorders. With our tools we have, for example, shown that protein aggregation causes a major problem for the system. We also found that an abnormal ubiquitin which has been found in Alzheimer’s disease interferes with the household functions of the ubiquitin/proteasome system. Finally, we demonstrated that cellular stress conditions, which are commonly found in neurodegenerative diseases, affects the functionality of the ubiquitin/proteasome system.

We have been trying to elucidate why the ubiquitin/proteasome system works less efficient during stress conditions. To our surprise, we found that probably the protein ubiquitin itself is the bottleneck. Despite the fact that high levels of ubiquitin are present in all cells, the levels are still rate limiting which is probably due to the fact that ubiquitylation is involved in so many cellular processes. As a consequence different ubiquitin-dependent processes are competing for a limited pool of ubiquitin and especially during stress when much more ubiquitin is required for degradation the system runs havoc. Based on this finding we postulated the ‘ubiquitin equilibrium hypothesis’ according to which the limited amount of ubiquitin enables crosstalk between various ubiquitin-dependent processes. We indeed found that blocking ubiquitin-dependent degradation directly affects ubiquitin-dependent transcription by competing ubiquitin from histones which dictate transcriptional activity. At the moment, we are also looking at other ubiquitin-dependent processes to get a better idea of the level of crosstalk and the relevance if this crosstalk in diseases.

Inspired by our earlier work in which we studied how certain proteins resist proteasomal degradation, we started to work on Rad23, a protein involved in nucleotide excision repair which protects our genome from UV-induced DNA damage. Interestingly, Rad23 has to interact with the proteasome when the DNA is being repaired but somehow it escapes from degradation. We identified the domain (stabilization signal) that is responsible for protecting Rad23 and showed that the stabilization signal of Rad23 is important for DNA repair. Thus, our research shows that some proteins protect themselves from degradation by the proteasome, a situation that we had only observed previously for pathologic proteins such as a viral protein and proteins causing neurodegenerative diseases. We also found that UV light causes mono-ubiquitylation of histones and that this is ubiquitylation is part of the DNA repair response, which is another example of the direct link between DNA repair and the ubiquitin/proteasome system. The functional significance of this DNA damage-induced histone modification remains unclear.

In our ongoing research, we focus on these fascinating aspects of the ubiquitin/proteasome system and are ready for new surprises.

Please visit our website for more information: Dantuma lab..

About our research:

'Ubiquitin Tug-O-War' by Rabiya S. Tuma. 2006. J. Cell Biol. 173(1): 3.

‘Första signalen som skyddar protein i cellen mot nedbrytning’ by Hanna Meerveld. 2005. BiotechSweden 2005(5): 5. (Swedish)

‘Waste disposal under the spotlight’ by M. Brazil. 2003. Nature Rev. Neurosc. 4:698.

‘Ubikvitin: Det lilla proteinet som styr livet’ by G. Strachal. 2001. Medicinsk Vetenskap 2001(4): 2-5. (Swedish)

Key references from the Dantuma lab:

Bergink, S., F.A. Salomons, D. Hoogstraten, T.A.M. Groothuis, , H. de Waard, J. Wu, L. Yuan, E. Citterio, A. Houtsmuller, J. Neefjes, J.H.J. Hoeijmakers, W. Vermeulen and N.P. Dantuma. 2006. DNA damage triggers nucleotide excision repair-dependent monoubiquitylation of histone H2A. Genes Dev. 20:1343-1352.

Dantuma, N.P., T.A.M. Groothuis, F.A. Salomons and J. Neefjes. 2006. A dynamic ubiquitin equilibrium couples proteasomal activity to chromatin remodeling. J. Cell Biol. 173: 19-26.

Heessen, S., M.G. Masucci and N.P. Dantuma. 2005. The UBA2 domain functions as an intrinsic stabilization signal that protects Rad23 from proteasomal degradation. Mol. Cell 18: 225-235.

Lindsten, K., V. Menéndez-Benito, M.G.Masucci and N.P. Dantuma. 2003. A transgenic mouse model of the ubiquitin-proteasome system. Nature Biotechnol. 21: 897-902.

Neefjes, J., and N.P. Dantuma. 2004. Fluorescent probes for proteolysis: tools for drug discovery. Nature Rev. Drug Discov. 3: 58-69.