DNA end-binding specificity of human Rad50/Mre11 is influenced by ATP.
The Rad50, Mre11 and Nbs1 complex is involved in many essential chromosomal organization processes dealing with DNA ends, including two major pathways of DNA double-strand break repair, homologous recombination and non-homologous end joining. Previous data on the structure of the human Rad50 and Mre11 (R/M) complex suggest that a common role for the protein complex in these processes is to provide a physical link between DNA ends such that they can be processed in an organized and coordinated manner. Here we describe the DNA binding properties of the R/M complex. The complex bound to both single-stranded and double-stranded DNA. Scanning force microscopy analysis of DNA binding by R/M showed the requirement for an end to form oligomeric R/M complexes, which could then migrate or transfer away from the end. The R/M complex had a lower preference for DNA substrates with 3'-overhangs compared with blunt ends or 5'-overhangs. Interestingly, ATP binding, but not hydrolysis, increased the preference of R/M binding to DNA substrates with 3'-overhangs relative to substrates with blunt ends and 5'-overhangs.
de Jager
Martijn
M
Department of Cell Biology and Genetics, Erasmus MC, PO Box 1738, 3000 DR Rotterdam, The Netherlands.
Wyman
Claire
C
van Gent
Dik C
DC
Kanaar
Roland
R
D017766:Acid Anhydride Hydrolases; D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D004247:DNA / Q000378:metabolism* / Q000648:ultrastructure; D045643:DNA Repair Enzymes; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D000076228:MRE11 Homologue Protein; D046911:Macromolecular Substances; D018625:Microscopy, Atomic Force; D011485:Protein Binding
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; C121766:MRE11 protein, human; D046911:Macromolecular Substances; D000255:Adenosine Triphosphate; D004247:DNA; D000076228:MRE11 Homologue Protein; D017766:Acid Anhydride Hydrolases; C112158:RAD50 protein, human; D045643:DNA Repair Enzymes