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PMID: 15226437 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The NEF4 complex regulates Rad4 levels and utilizes Snf2/Swi2-related ATPase activity for nucleotide excision repair.

Molecular and cellular biology ·Vol. 24 ·No. 14 ·2004-07-00 ·Pages 6362-78

Ramsey KL, Smith JJ, Dasgupta A, Maqani N, Grant P, Auble DT

Abstract

Nucleotide excision repair factor 4 (NEF4) is required for repair of nontranscribed DNA in Saccharomyces cerevisiae. Rad7 and the Snf2/Swi2-related ATPase Rad16 are NEF4 subunits. We report previously unrecognized similarity between Rad7 and F-box proteins. Rad16 contains a RING domain embedded within its ATPase domain, and the presence of these motifs in NEF4 suggested that NEF4 functions as both an ATPase and an E3 ubiquitin ligase. Mutational analysis provides strong support for this model. The Rad16 ATPase is important for NEF4 function in vivo, and genetic analysis uncovered new interactions between NEF4 and Rad23, a repair factor that links repair to proteasome function. Elc1 is the yeast homologue of a mammalian E3 subunit, and it is a novel component of NEF4. Moreover, the E2s Ubc9 and Ubc13 were linked to the NEF4 repair pathway by genetic criteria. Mutations in NEF4 or Ubc13 result in elevated levels of the DNA damage recognition protein Rad4 and an increase in ubiquitylated species of Rad23. As Rad23 also controls Rad4 levels, these results suggest a complex system for globally regulating repair activity in vivo by controlling turnover of Rad4.

MeSH Terms
Adenosine Triphosphatases/genetics,metabolism Amino Acid Sequence DNA Damage DNA Repair DNA-Binding Proteins/genetics,metabolism Fungal Proteins/genetics,metabolism Macromolecular Substances Molecular Sequence Data Phenotype Protein Structure, Tertiary Protein Subunits/genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism,radiation effects Saccharomyces cerevisiae Proteins/genetics,metabolism Ultraviolet Rays
Chemicals
DNA-Binding Proteins Fungal Proteins Macromolecular Substances Protein Subunits RAD23 protein, S cerevisiae RAD7 protein, S cerevisiae Saccharomyces cerevisiae Proteins Adenosine Triphosphatases RAD16 protein, S cerevisiae
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ramsey Kerrington L
Department of Biochemistry and Molecular Genetics, University of Virginia Health System, Charlottesville, VA 22908-0733, USA.
Smith Joshua J
Dasgupta Arindam
Maqani Nazif
Grant Patrick
Auble David T
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.24.14.6362-6378.2004
Published
2004-07-00
Pages
6362-78
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC434245
Subset
IM
Grants
NIGMS NIH HHS · R01 GM055763 · United States
NIGMS NIH HHS · GM55763 · United States
External Links
PubMed source DOI: 10.1128/MCB.24.14.6362-6378.2004 PMC Full Text View journal details
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