In vivo Interaktion von Proteinen der Kernhülle In vivo interactions of nuclear envelope proteins

• Applicant: Professor Dr. Georg Krohne
• Subject Area: Biochemistry
• Term: from 2000 to 2007
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 5203594

Integral and peripheral proteins of the inner nuclear membrane interact with each other, with proteins of the nuclear lamina, and components of the peripheral chromatin. In vivo and in vitro I shall investigate with three different strategies the structural and functional alterations of the cell nucleus when these interactions are disturbed. First, competetive peptides comprising sequences of binding domains of distinct nuclear envelope proteins will be expressed in tissue culture cells. In parallel the purified peptides will be microinjected into mitotic cells to analyze their function during nuclear reconstitution. Second, these peptides will be coupled to columns to enable the purification of binding partners from interphase and mitotic extracts. Third, the mRNA of one or more nuclear envelope proteins will be depleted by the microinjection of double stranded RNA (dsRNA = annealed sense and anti-sense RNA of a cDNA coding for a distinct nuclear envelope protein) into cultured cells, and early embryos of Xenopus and Drosophila. As a consequence of the mRNA depletion, after few cell divisions nuclei will contain very low amounts of the specific nuclear envelope protein.

• DFG Programme: Priority Programmes
• Subproject of: SPP 1050: Funktionelle Architektur des Zellkerns