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Overexpression of wild‐type and mutant NDP kinase in Dictyostelium discoideum
Author(s) -
Sellam Olivier,
Véron Michel,
Hildebrandt Martin
Publication year - 1995
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1995.tb02393.x
Subject(s) - dictyostelium discoideum , biology , mutant , kinase , wild type , gene , microbiology and biotechnology , map3k7 , phosphoglycerate kinase , mutation , biochemistry , cyclin dependent kinase 2 , protein kinase a
Summary Nucleoside diphosphate (NDP) kinase has a central role in the synthesis of (deoxy‐)trinucleotides. In addition, mutations in the gene encoding NDP kinase have been shown to have important consequences for Drosophila development and mammalian tumorogenesis. We have overexpressed, in Dictyostelium discoideum , a genomic clone encoding the enzyme NDP kinase. The concomitant increase in the levels of RNA and enzyme activity identifies a 5′ non‐coding genomic region of 0.9 kb as being the complete promoter region. Overexpression of wild‐type NDP kinase has no effect on development. This is also true for an inactive mutant H122C that does not have a dominant inhibitor effect. Overexpression of the P105G mutant NDP kinase, which is known to be affected in its stability in vitro , only leads to a small increase in total NDP‐kinase activity. Thermal and chemical denaturation experiments demonstrate the formation of hexameric hybrids between wild‐type and mutant monomers.