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PMID: 12411510 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The flexible loop of human FEN1 endonuclease is required for flap cleavage during DNA replication and repair.

The EMBO journal ·Vol. 21 ·No. 21 ·2002-11-01 ·Pages 5930-42

Storici F, Henneke G, Ferrari E, Gordenin DA, Hübscher U, Resnick MA

Abstract

The conserved, structure-specific flap endonuclease FEN1 cleaves 5' DNA flaps that arise during replication or repair. To address in vivo mechanisms of flap cleavage, we developed a screen for human FEN1 mutants that are toxic when expressed in yeast. Two targets were revealed: the flexible loop domain and the catalytic site. Toxic mutants caused G(2) arrest and cell death and were unable to repair methyl methanesulfonate lesions. All the mutant proteins retained flap binding. Unlike the catalytic site mutants, which lacked cleavage of any 5' flaps, the loop mutants exhibited partial ability to cut 5' flaps when an adjacent single nucleotide 3' flap was present. We suggest that the flexible loop is important for efficient cleavage through positioning the 5' flap and the catalytic site.

MeSH Terms
Base Sequence Catalytic Domain DNA DNA Repair/physiology DNA Replication/physiology Endodeoxyribonucleases/chemistry,genetics,metabolism,physiology Exodeoxyribonuclease V Exodeoxyribonucleases/chemistry,genetics,metabolism,physiology Flap Endonucleases Humans Hydrolysis Models, Molecular Mutation
Chemicals
DNA Endodeoxyribonucleases Exodeoxyribonucleases Flap Endonucleases FEN1 protein, human Exodeoxyribonuclease V
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Storici Francesca
Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.
Henneke Ghislaine
Ferrari Elena
Gordenin Dmitry A
Hübscher Ulrich
Resnick Michael A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/cdf587
Published
2002-11-01
Pages
5930-42
Language
English
Region
England
NLM ID
8208664
PMCID
PMC131084
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/cdf587 PMC Full Text View journal details
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