Characterisation of the SAGA/ADA complex in Aspergillus nidulans by tandem affinity purification

Ref ID: 18308

Author:

Fischer, J., Nuetzmann, H.-W., Schroeckh, V. and Brakhage

Author address:

, A. A. Department Molecular and Applied Microbiology, LeibnizInstitute for Natural Product Research and Infection Biology (HKI), Jena, Germany, and Institute of Microbiology, Friedrich Schiller
University Jena, Germany Juliane.Fischer@hki-jena.de

Full conference title:

Asperfest 9

Abstract:

The recent finding that the histone acetyltransferase (HAT) complex SAGA/ADA mediates the response of the fungus Aspergillus
nidulans to the bacterium Streptomyces rapamycinicus (1) opens up a number of questions. It was shown that the SAGA/ADA
complex is involved in the regulation of the orsellenic/ lecanoric acid biosynthesis gene cluster, as deletion of its HAT-encoding gene
gcnE resulted in the lack of orsA transcription during co-cultivation. In order to investigate the SAGA/ADA complex in A. nidulans,
the complex subunits GcnE and AdaB were tagged and purified by tandem affinity purification (TAP). This method is especially
suited for the purification of protein complexes under native conditions. Therefore, the TAP-tag method represents an appropriate
system for the investigation and analysis of the SAGA/ADA complex composition under various conditions. The TAP-tag constructs
were assembled by fusion PCR and transformed directly into A. nidulans via homologues recombination. Western blotting was
performed to monitor the purification procedure. For AdaB-TAP and GcnE-TAP bands of the expected size were detected. However,
further optimisation of the purification procedure is required. (1) Nuetzmann et al. (2011) PNAS

Abstract Number: 59)

Conference Year: 2012

Link to conference website: NULL

New link: NULL

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