Establishment of sister chromatid cohesion at the S. cerevisiae replication fork.
Two identical sister copies of eukaryotic chromosomes are synthesized during S phase. To facilitate their recognition as pairs for segregation in mitosis, sister chromatids are held together from their synthesis onward by the chromosomal cohesin complex. Replication fork progression is thought to be coupled to establishment of sister chromatid cohesion, facilitating identification of replication products, but evidence for this has remained circumstantial. Here we show that three proteins required for sister chromatid cohesion, Eco1, Ctf4, and Ctf18, are found at, and Ctf4 travels along chromosomes with, replication forks. The ring-shaped cohesin complex is loaded onto chromosomes before S phase in an ATP hydrolysis-dependent reaction. Cohesion establishment during DNA replication follows without further cohesin recruitment and without need for cohesin to re-engage an ATP hydrolysis motif that is critical for its initial DNA binding. This provides evidence for cohesion establishment in the context of replication forks and imposes constraints on the mechanism involved.
Lengronne
Armelle
A
Chromosome Segregation Laboratory, Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, 44 Lincoln's Inn Fields, London WC2A 3PX, United Kingdom.
McIntyre
John
J
Katou
Yuki
Y
Kanoh
Yutaka
Y
Hopfner
Karl-Peter
KP
Shirahige
Katsuhiko
K
Uhlmann
Frank
F
D000123:Acetyltransferases / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism; D020816:Amino Acid Motifs / Q000502:physiology; D018797:Cell Cycle Proteins / Q000032:analysis / Q000378:metabolism; D002842:Chromatids / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis / Q000378:metabolism; D004261:DNA Replication / Q000502:physiology*; D004271:DNA, Fungal / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D008957:Models, Genetic; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D016196:S Phase / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000378:metabolism / Q000502:physiology; D000097722:Cohesins
D000123:Acetyltransferases; D000251:Adenosine Triphosphatases; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D029701:Saccharomyces cerevisiae Proteins; D000097722:Cohesins; C092693:CTF18 protein, S cerevisiae; C078338:CTF4 protein, S cerevisiae; C118076:ECO1 protein, S cerevisiae