The Svedberg Award 2006

Piergiorgio Percipalle
Department of Cell and Molecular Biology, Karolinska Institutet

Background
After graduating in Chemistry, I started my PhD at the International Centre for Genetic Engineering and Biotechnology, Trieste, where I worked on the molecular mechanisms of protein-DNA recognition using chemical, biochemical and biophysical assays. I obtained my PhD in molecular genetics at the International School for Advanced Studies with a thesis entitled “Engineering DNA-binding proteins based on the helix-turn-helix motif”. I then moved to the MRC Laboratory of Molecular Biology, Cambridge, where I did a postdoc in the laboratory of Daniela Rhodes. In Daniela’s lab I mapped some of the protein-protein interactions that take place during protein import into the cell nucleus. That period was very creative for me since I started thinking about why actin is imported into the cell nucleus where it is such an abundant protein (about 15% of cellular actin is present in the cell nucleus). I then moved to the Karolinska Institutet in the laboratory of Bertil Daneholt where I became acquainted with the polytene chromosomes of the dipteran insect Chironomus tentans, ideal to study gene expression in situ. In that period, the polytene chromosomes turned out to be instrumental to establish that actin is in active transcription sites and it is a genuine component of nascent pre-messenger-ribonucleoprotein (pre-mRNP) particles. In January 2004, I became a principal investigator in the Department of Cell and Molecular Biology (CMB) at the Karolinska Institutet. Since then, we have shown that actin is an essential transcription factor in both RNA polymerase I (pol I) and RNA polymerase II (pol II)-mediated transcription.

Research
The research in my group focuses on the mechanisms of transcriptional and post-transcriptional control of gene expression. We are studying both normal cells and cancer cells where these mechanisms are profoundly affected and deregulated, leading to disruption of controlled gene activity and malignant cell transformation.
One of our main interests is to understand the crosstalk between transcription apparatus and dynamic alterations of chromatin structure which are mediated by chromatin remodelling complexes and histone modifying enzymes. These factors are recruited on active genes through complex relay mechanisms which are not yet fully understood. Several independent studies have shown that nuclear actin may perform a pivotal role in this dynamic recruitment since it is present in certain ATP-dependent chromatin remodelling complexes, in pre-mRNP particles and it is associated with all three eukaryotic RNA polymerases. Some mechanistic insights came from the discovery of nuclear actin-binding proteins. In ribosomal gene transcription, the interaction between actin and nuclear myosin 1 (NM1) is required to recruit the novel chromatin remodelling complex B-WICH, an activator of pol I transcription elongation. On the other hand, pol II transcription requires the interaction between actin and the heterogeneous ribonucleoprotein hnRNP U. This mechanism appears to be conserved from humans to insects and is likely to be required for the recruitment of specific histone acetyl transferases to maintain a productive pol II transcription elongation phase. Based on the above observations, we have proposed that actin functions in molecular switches as an allosteric regulator of RNA polymerase-mediated transcription. Molecular switches may represent a means of recruiting co-factors on active genes, with the help of specific adaptors, NM1 in pol I transcription and hnRNP U in pol II transcription.
This model raises many intriguing questions. We are currently focusing on two of them, namely “what is the polymerization state of actin during its function along actively transcribed genes?” and “are actin-based molecular switches required only for transcription or they are also utilized down-stream for post-transcriptional control of gene expression?

Selected references
Percipalle P * (2007) Genetic connections of the actin cytoskeleton and beyond. BioEssays,
in press

Percipalle P *, Östlund Farrants A-K (2006) Chromatin remodelling and transcription:
Be-WICHed by nuclear myosin 1. Current Opinion Cell Biology, 18: 267-274
Percipalle P *, Visa N (2006) Molecular functions of nuclear actin. J Cell Biol, 172: 967-971
Percipalle P *, Fomproix N, Cavellan E, Voit R, Reimer G, Krüger T, Thyberg J, Scheer U, Grummt I, Östlund Farrants A-K (2006) The chromatin remodelling complex WSTF-SNF2h interacts with nuclear myosin 1and serves a role in RNA polymerase I transcription.
EMBO Reports
7: 525-530
Kukalev A, Nord Y, Palmberg C, Bergman T, Percipalle P * (2005) Actin and hnRNP U cooperate for productive transcription by RNA polymerase II. 2004. Nature Struct Mol Biol 12: 238-244
Percipalle P, Fomproix N, Kylberg K, Miralles M, Björkroth B, Daneholt B, Visa N (2003) An actin-ribonucleoprotein interaction is involved in transcription by RNA polymerase II. Proc. Natl Acad Sci USA 100: 6475-6480