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Titill

Chromatin remodeling complexes in the Yeast Schizosaccharomyces pombe

Útdráttur

In eukaryotic cells, an important part of gene regulation is the accessibility of genes to transcription. The chromatin needs to be modified for genes to become accessible. Chemical groups can be added or removed from the histone proteins but modifications also take place by nucleosomal sliding or histone exchange. These alterations are controlled by chromatin modifying enzymes, which are often parts of big complexes, called chromatin remodeling complexes. Accessibility also regulates other processes involving DNA, such as DNA replication and repair. It has recently been demonstrated that some chromatin remodeling complexes, notably the INO80, SWR1/ SRCAP and NuA4/TIP60, play an important role in DNA double-strand break repair and checkpoint responses. Many chromatin remodeling complexes have been characterized in different organisms, such as human, the fruit fly and the yeast Saccharomyces cerevisiae. For some reason, work on chromatin remodeling in the important model organism Schizosaccharomyces pombe has been lagging. S. pombe is distantly related to S. cerevisiae and has proven to be in many ways a suitable model organism to study the eukaryotic core pathways of DNA metabolism and cell cycle regulation. The aim of the project was to characterize chromatin remodeling complexes in the yeast S. pombe. Focusing on three important complexes; INO80, SWR1 and NuA4, we have epitope tagged the proteins Ino80, Swr1, Rvb1, Rvb2 and Epl1 which have all been implicated in chromatin remodeling in other organisms. After tagging the proteins, we isolated the co-purifying complexes using immunoprecipitation. The subunit composition of the complexes was then characterized by multidimensional tandem mass spectrometry (2D-LC-MS/MS-MudPit).

Birting
7.7.2010


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