title: Domain architecture of the DRpp29 protein and its interaction with the RNA subunit of Dictyostelium discoideum RNase P.
abstract: Dictyostelium discoideum nuclear RNase P is a ribonucleoprotein complex that displays similarities with its counterparts from higher eukaryotes such as the human enzyme, but at the same time it retains distinctive characteristics. In the present study, we report the molecular cloning and interaction details of DRpp29 and RNase P RNA, two subunits of the RNase P holoenzyme from D. discoideum. Electrophoretic mobility shift assays exhibited that DRpp29 binds specifically to the RNase P RNA subunit, a feature that was further confirmed by the molecular modeling of the DRpp29 structure. Moreover, deletion mutants of DRpp29 were constructed in order to investigate the domains of DRpp29 that contribute to and/or are responsible for the direct interaction with the D. discoideum RNase P RNA. A eukaryotic specific, lysine- and arginine-rich region was revealed, which seems to facilitate the interaction between these two subunits. Furthermore, we tested the ability of wild-type and mutant DRpp29 to form active RNase P enzymatic particles with the Escherichia coli RNase P RNA.
authors.affiliation: Department of Biochemistry, School of Medicine, University of Patras, 26500 Patras, Greece.
authors.forename: Vassiliki
authors.initials: V
authors.lastname: Stamatopoulou
authors.forename: Chrisavgi
authors.initials: C
authors.lastname: Toumpeki
authors.forename: Andreas
authors.initials: A
authors.lastname: Tzakos
authors.forename: Anastassios
authors.initials: A
authors.lastname: Vourekas
authors.forename: Denis
authors.initials: D
authors.lastname: Drainas
mesh_terms: D015152:Blotting, Northern; D004023:Dictyostelium / Q000201:enzymology*; D024202:Electrophoretic Mobility Shift Assay; D047468:Immunoprecipitation; D009154:Mutation; D011485:Protein Binding; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D043262:Ribonuclease P / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D021122:Protein Subunits; D016337:RNA, Catalytic; D043262:Ribonuclease P