Storici, F; Henneke, G; Ferrari, E; Gordenin, D A; Hübscher, U; Resnick, M A (2002). The flexible loop of human FEN1 endonuclease is required for flap cleavage during DNA replication and repair. The EMBO Journal, 21(21):5930-5942.
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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.
|Item Type:||Journal Article, refereed|
|Communities & Collections:||05 Vetsuisse Faculty > Institute of Veterinary Biochemistry and Molecular Biology|
|Dewey Decimal Classification:||570 Life sciences; biology|
|Date:||1 November 2002|
|Deposited On:||11 Feb 2008 12:18|
|Last Modified:||27 Nov 2013 18:05|
|Publisher:||European Molecular Biology Organization ; Nature Publishing Group|
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