Screen for PI3P effectors of autophagy
Class III PI3 kinase and its product PI3P are essential for autophagy. We have performed an siRNA screen targeting putative phosphoinositide-binding proteins in mammalian cells and in Drosophila melanogaster and will analyse screen candidates for a function in autophagy. This project involves a collaboration with Thomas Neufeld, University of Minnesota and is funded by The Norwegian Centennial Chair program.
Functional characterization of the autophagic adapter protein Alfy
The large PI3P-binding protein Alfy (autophagy linked FYVE protein) is recruited to intracellular protein aggregates and is required to target such aggregates for degradation by autophagy (Filimonenko et al., MolCell 2010). We will elucidate the role of the various functional domains in Alfy in order to understand its function in protein aggregate formation and degradation.
The molecular mechanisms of aggrephagy
We will use siRNA screens and targeted pulldown assays to identify new players involved in protein aggregate-mediated autophagy (aggrephagy).
The role of autophagy in degradation of nuclear waste
Although autophagy is a strictly cytoplasmic process, we have evidence that autophagy also plays a role in deposition of nuclear “waste products” such as misfolded proteins and damaged DNA. We are currently in the process of investigating the mechanisms involved and how this is regulated.
The role of autophagy in degradation of leukemia-associated oncoproteins
We have recently found that autophagy mediates degradation of the acute promyelocytic leukemia (APL) oncoprotein PML-RARA and that drugs known to cause clinical remission in APL patients induce autophagy and PML-RARA degradation (Isakson et al, Blood 2010). We will investigate whether autophagy also plays a role in clearance of other leukemia-associated fusion proteins.