Yeast resolving enzyme CCE1 makes sequential cleavages in DNA junctions within the lifetime of the complex.

TY - JOUR

T1 - Yeast resolving enzyme CCE1 makes sequential cleavages in DNA junctions within the lifetime of the complex.

AU - Fogg, JM

AU - Schofield, MJ

AU - Déclais, AC

AU - Lilley, DM

PY - 2000/4

Y1 - 2000/4

N2 - CCE1 is a DNA junction-resolving enzyme of Saccharomyces cerevisiae. Such enzymes are required to make two symmetrically paired cleavages in order to resolve the four-way junction productively. Using a cruciform assay, we show here that CCE1 introduces two unilateral cleavages in a sequential manner. This requires that the protein remains bound to the junction, preventing branch migration of the point of strand exchange. From a detailed kinetic analysis, we find that the CCE1 cleavage at a given site is accelerated by a factor of 5-10 when it occurs subsequently to the initial cleavage. These properties ensure a productive resolution of the four-way junction and may be general for junction-resolving enzymes.

AB - CCE1 is a DNA junction-resolving enzyme of Saccharomyces cerevisiae. Such enzymes are required to make two symmetrically paired cleavages in order to resolve the four-way junction productively. Using a cruciform assay, we show here that CCE1 introduces two unilateral cleavages in a sequential manner. This requires that the protein remains bound to the junction, preventing branch migration of the point of strand exchange. From a detailed kinetic analysis, we find that the CCE1 cleavage at a given site is accelerated by a factor of 5-10 when it occurs subsequently to the initial cleavage. These properties ensure a productive resolution of the four-way junction and may be general for junction-resolving enzymes.

UR - http://europepmc.org/abstract/med/10747798

U2 - 10.1021/bi992785v

DO - 10.1021/bi992785v

M3 - Article

C2 - 10747798

SN - 0006-2960

VL - 39

SP - 4082

EP - 4089

JO - Biochemistry

JF - Biochemistry

IS - 14

ER -