International Network of Plant Abiotic Stress

COST STSM Reference Number: COST-STSM-FA0605-5189

STSM Grantee: Ms Mary Prathiba Joseph, Biological Research Center, Szeged (HU), mary.prathiba@gmail.com

Host Institute: Csaba Koncz, Max-Planck-Institute für Züchtungsforschung,D-50829 Köln (DE), koncz@mpiz-koeln.mpg.de

Period: 19/10/2009 to 19/11/2009

I  chose to work at the MPI for Plant Breeding Research, Cologne, Germany  for the STSM program under the able guidance of Prof.Csaba Koncz and Dr. Femke De Jong (Post Doc Fellow).  Our aim was to identify protein complexes and protein interacting partners of a previously unknown Zn finger transcription factor (CAI1) which regulates ABA responses in Arabidopsis.

The activity involved training in the following biochemical technologies:

Ø    Nuclear protein extraction from seedlings.

Ø    Ni-NTA column purification under native conditions.

Ø    TCA precipitation and Mass spec result analysis.

We were successful in optimizing the nuclear protein isolation from seedlings by the protocol completely standardized by the host laboratory (Prof. Csaba Koncz & Dr. Femke De Jong). We could also optimize the affinity column purification of the CAI1 protein and also co-purify several other proteins. Mass Spectrometry analysis  revealed the presence of a number of proteins in the purified samples.  Analysis of control samples have not been completed by the end of the fellowship, therefore the list of the CAI1 interacting proteins cannot be established yet.

However, repetition of the experiments with more stringent conditions of elution of CAI1 from the nickel affinity matrix is necessary to further minimize the contamination of non specific proteins, and to confirm the co-purified proteins as CAI1-binding partners, is essential.

Due to succesful training of the applicant, further studies and repetition of the experiments with appropriate controls will be done in the home laboratory at the BRC, Szeged, Hungary.

Collaboration between the home and host laboratories is foreseen and will include further analysis of the interaction of CAI1 protein and newly identified nuclear factors. The grantee, Mary Prathiba Joseph will maintain close information exchange with the host laboratory, which will supply data about control samples, and will advice in the analysis and confirmation of protein-protein interactions.