title: AtRAD5A is a DNA translocase harboring a HIRAN domain which confers binding to branched DNA structures and is required for DNA repair in vivo.
abstract: DNA lesions such as crosslinks represent obstacles for the replication machinery. Nonetheless, replication can proceed via the DNA damage tolerance pathway also known as postreplicative repair pathway. SNF2 ATPase Rad5 homologs, such as RAD5A of the model plant Arabidopsis thaliana, are important for the error-free mode of this pathway. We able to demonstrate before, that RAD5A is a key factor in the repair of DNA crosslinks in Arabidopsis. Here, we show by in vitro analysis that AtRAD5A protein is a DNA translocase able to catalyse fork regression. Interestingly, replication forks with a gap in the leading strand are processed best, in line with its suggested function. Furthermore AtRAD5A catalyses branch migration of a Holliday junction and is furthermore not impaired by the DNA binding of a model protein, which is indicative of its ability to displace other proteins. Rad5 homologs possess HIRAN (Hip116, Rad5; N-terminal) domains. By biochemical analysis we were able to demonstrate that the HIRAN domain variant from Arabidopsis RAD5A mediates structure selective DNA binding without the necessity for a free 3'OH group as has been shown to be required for binding of HIRAN domains in a mammalian RAD5 homolog. The biological importance of the HIRAN domain in AtRAD5A is demonstrated by our result that it is required for its function in DNA crosslink repair in vivo.
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Kobbe
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Andy
authors.initials: A
authors.lastname: Kahles
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Walter
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Klemm
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Mannuss
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Knoll
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Focke
authors.affiliation: Botanical Institute II, Karlsruhe Institute of Technology (KIT), Karlsruhe, 76131, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Puchta
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004249:DNA Damage / Q000235:genetics / Q000502:physiology; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair / Q000235:genetics / Q000502:physiology; D004261:DNA Replication / Q000235:genetics / Q000502:physiology; D011485:Protein Binding
chemical_list: D029681:Arabidopsis Proteins; C561786:RAD5a protein, Arabidopsis; D004247:DNA; D004265:DNA Helicases
keywords: 
  Arabidopsis thaliana
  ; At5g22750; DNA crosslink repair; DNA damage tolerance; DNA translocase; DNA-binding domain; HIRAN domain; Rad5; postreplicative repair; replication fork