Reference: Bailly V, et al. (1997) Yeast DNA repair proteins Rad6 and Rad18 form a heterodimer that has ubiquitin conjugating, DNA binding, and ATP hydrolytic activities. J Biol Chem 272(37):23360-5

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Abstract


The RAD6 and RAD18 genes of Saccharomyces cerevisiae are required for postreplicative bypass of ultraviolet (UV)-damaged DNA and for UV mutagenesis. The RAD6 encoded protein is a ubiquitin conjugating enzyme, and RAD18 encodes a protein containing a RING finger motif and a nucleotide binding motif. Rad18 can be co-immunoprecipitated with Rad6, indicating that the two proteins exist in a complex in vivo. Here, we co-overproduce the two proteins using a yeast multicopy plasmid, purify the Rad6-Rad18 complex to near homogeneity, and show that the complex is heterodimeric. The Rad6-Rad18 heterodimer has ubiquitin conjugating activity, binds single-stranded DNA, and possesses single-stranded DNA-dependent ATPase activity. The Rad6-Rad18 complex provides the first example wherein a ubiquitin conjugating activity is physically associated with DNA binding and ATPase activities provided by an associated protein factor. The co-existence of these activities should provide the complex with the ability to recognize single-stranded DNA resulting from stalling of the replication machinery at DNA damage sites and to recognize the components of the DNA replication machinery for ubiquitination by Rad6.

Reference Type
Journal Article | Research Support, U.S. Gov't, P.H.S.
Authors
Bailly V, Lauder S, Prakash S, Prakash L
Primary Lit For
RAD18 | RAD6 | RAD6-RAD18 ubiquitin ligase complex
Additional Lit For
RAD6-UBR2 ubiquitin ligase complex | MUB1-RAD6-UBR2 ubiquitin ligase complex

Gene Ontology Annotations 11 entries for 3 genes


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Gene/ComplexQualifierGene Ontology TermAnnotation ExtensionEvidenceSourceAssigned On
RAD6-RAD18 ubiquitin ligase complexinvolved inerror-free postreplication DNA repairBSRComplexPortal2018-03-09
RAD6-RAD18 ubiquitin ligase complexenablesATP bindingIPIComplexPortal2018-03-09
RAD6-RAD18 ubiquitin ligase complexenablesubiquitin-protein transferase activityIDAComplexPortal2018-03-09
RAD6-RAD18 ubiquitin ligase complexinvolved inprotein monoubiquitinationIDAComplexPortal2018-03-09
RAD18contributes tosingle-stranded DNA bindinghas input single-stranded DNAIDASGD2019-12-09
RAD18part ofRad6-Rad18 complexIDASGD2013-10-11
RAD18contributes tosingle-stranded DNA helicase activityIDASGD2019-12-09
RAD6enablesubiquitin-protein transferase activityIDASGD2013-08-07
RAD6part ofRad6-Rad18 complexIDASGD2013-10-11
RAD6contributes tosingle-stranded DNA bindinghas input single-stranded DNAIDASGD2019-12-09
Showing 1 to 10 of 11 entries

Interaction Annotations


Genetic Interactions

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 5 entries for 4 genes

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InteractorInteractorAssayAnnotationActionModification
RAD18RAD6Affinity Capture-Westernmanually curatedHit-BaitNo Modification

bound in the absence or presence of single stranded DNA

RAD18RAD6Reconstituted Complexmanually curatedHit-BaitNo Modification
RAD18RAD6Co-purificationmanually curatedHit-BaitNo Modification

Rad6 formed a thioester adduct with GST-ubiquitin in an in vitro assay with purified Uba1, Rad6, ATP and GST-ub; source of ubiquitin is unknown

UBA1RAD6Biochemical Activitymanually curatedBait-Hitubiquitinylated lysine

likely ubiquitin conjugate

UBA4RAD6Reconstituted Complexmanually curatedHit-BaitNo Modification
Showing 1 to 5 of 5 entries