Nucleic acid-binding properties of the RRM-containing protein RDM1.
RDM1 (RAD52 Motif 1) is a vertebrate protein involved in the cellular response to the anti-cancer drug cisplatin. In addition to an RNA recognition motif, RDM1 contains a small amino acid motif, named RD motif, which it shares with the recombination and repair protein, RAD52. RDM1 binds to single- and double-stranded DNA, and recognizes DNA distortions induced by cisplatin adducts in vitro. Here, we have performed an in-depth analysis of the nucleic acid-binding properties of RDM1 using gel-shift assays and electron microscopy. We show that RDM1 possesses acidic pH-dependent DNA-binding activity and that it binds RNA as well as DNA, and we present evidence from competition gel-shift experiments that RDM1 may be capable of discrimination between the two nucleic acids. Based on reported studies of RAD52, we have generated an RDM1 variant mutated in its RD motif. We find that the L119GF --> AAA mutation affects the mode of RDM1 binding to single-stranded DNA.
Hamimes
Samia
S
International Agency for Research on Cancer (IARC), 150 Cours Albert Thomas, 69372 Lyon, France.
Bourgeon
Dominique
D
Stasiak
Alicja Z
AZ
Stasiak
Andrzej
A
Van Dyck
Eric
E
D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D002645:Chickens; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics* / Q000648:ultrastructure; D024202:Electrophoretic Mobility Shift Assay; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009154:Mutation; D012313:RNA / Q000737:chemistry* / Q000648:ultrastructure; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics* / Q000648:ultrastructure
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D016601:RNA-Binding Proteins; D012313:RNA