Interaktionen des testikulären outer dense fibre protein 2 (ODF 2)bei der zellulären Organisation

Interactions of the testicular outer dense fibre protein 2 (ODF 2) during cellular organisation

by Fatima Donkor
Doctoral thesis
Date of Examination:2006-10-31
Date of issue:2008-02-18
Advisor:PD Dr. Sigrid Hoyer-Fender
Referee:Prof. Dr. W. Engel
Referee:Prof. Dr. Ivo Feussner
Referee:Prof. Dr. Ulrich Grossbach
Persistent Address: http://dx.doi.org/10.53846/goediss-3391

 

 

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Abstract

English

Outer dense fibre 2 was initially identified as a main component of the sperm tail cy-toskeleton and was shown as a self-interacting centrosomal protein in somatic cells with affinity for microtubules. ODF2 has different localization in somatic versus male germ cells suggesting the possi-bility of different function. It was an aim to understand how the testicular protein ODF2 is involved in the cellular organisation of somatic cells and at the same time to identify puta-tive interaction partners which eventually take part in this process. A yeast two hybrid screen enabled to identify putative ODF2 interaction partners which are expected to be involved in protein transport, microtubuli binding and apoptotic proc-esses. The known ODF2 interactive partner ODF1 (Shao et al., 1997) co localized in somatic cells with centrosomal structures. Odf2/Odf1 cellular co-transfection experiments demonstrate a fibrillar molecular protein network generated by ODF1-ODF2 interaction event. ODF2 and dynamitin-overexpressing cells indicated an involvement of ODF2 in dynein-dependent protein assembly. ODF2 associated in somatic cells with the endoplasmtic reticulum and the Golgi complex. Immuncytology experiments with truncated ODF2 GFP fusion proteins suggested that the C terminal domain of ODF2 is thereby involved in binding to these specific organelles. Time series incubation of cells in culture with Brefeldin A (BFA) to analyse intracellular protein transport on ODF2 overexpressing cells were performed. Cells lead to similar but not identical results. Thus, the effects of short-term treatment with BFA was different from those of long-term treatment. Proliferation assays and flow cytometry analyses demonstrated that ODF2 overexpress-ing cells lead to a significant decrease of proliferation by “losing
Keywords: Outer dense fibre protein; centrosom; ODF2; ODF1
Schlagwörter: Outer dense fibre Proteine; Centrosom; ODF2; ODF1
 
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