MitoCheck Project

Introduction

Cell division or Mitosis is one of the fundamental processes of life. It is of vital importance for cells to make sure everything goes right every time they divide. Mistakes during cell division contribute to cancer and, if occurred during reproduction, are a leading cause of infertility and mental retardation. However, it is still poorly understood how cells coordinate the many disparate but inter-locking processes during mitosis. One of the well established facts is that protein kinases - a certain type of enzymes - play a major role. Their importance has even been recognized by a Nobel Prize in 2001. The question that remains is exactly which molecules are altered by those kinases and how.

The aims of the project are to systematically hunt for genes and proteins crucial for mitosis, to defi ne how these proteins are changed by phosphorylation - which is what kinases do - and to assess whether mitotic kinases have any diagnostic or therapeutic potential in cancer treatment. Along the way, it will be necessary to develop advanced technologies to address those questions in an effi cient and highly automated way.

High through-put RNAi screening

MitoCheck is using RNA interference (RNAi) screens to identify all proteins that are required for mitosis in human cells, affinity purification and mass spectrometry to identify protein complexes and mitosis-specific phosphorylation sites on these, and small molecule inhibitors to determine which protein kinase is required for the phosphorylation of which substrate. MitoCheck is furthermore establishing clinical assays to validate mitotic proteins as prognostic biomarkers for cancer therapy. (Neumann, etc, Nature Methods, Vol.3, No.5, 385 - 390, 2006)

Image analysis and phenotype classification

Examples of phenotype classes out of movies. (click to enlarge the images below)

Movie examples

Click below to see the movie (.MOV file, 635KB, 640 x 480 pixels)