Cohesin interaction with centromeric minichromosomes shows a multi-complex rod-shaped structure.
Cohesin is the protein complex responsible for maintaining sister chromatid cohesion. Cohesin interacts with centromeres and specific loci along chromosome arms known as Chromosome Attachment Regions (CARs). The cohesin holocomplex contains four subunits. Two of them, Smc1p (Structural maintenance of chromosome 1 protein) and Smc3p, are long coiled-coil proteins, which heterodimerize with each other at one end. They are joined together at the other end by a third subunit, Scc1p, which also binds to the fourth subunit, Scc3p. How cohesin interacts with chromosomes is not known, although several models have been proposed, in part on the basis of in vitro assembly of purified cohesin proteins. To be able to observe in vivo cohesin-chromatin interactions, we have modified a Minichromosome Affinity Purification (MAP) method to isolate a CAR-containing centromeric minichromosome attached to in vivo assembled cohesin. Transmission Electron Microscopy (TEM) analysis of these minichromosomes suggests that cohesin assumes a rod shape and interacts with replicated minichromosome at one end of that rod. Additionally, our data implies that more than one cohesin molecule interacts with each pair of replicated minichromsomes. These molecules seem to be packed into a single thick rod, suggesting that the Smc1p and Smc3p subunits may interact extensively.
The Intercollege Graduate Program in Cell and Developmental Biology, The Huck Institutes of the Life Sciences, Pennsylvania State University, University Park, Pennsylvania, United States of America.
Alexandra
A
Surcel
Douglas
D
Koshland
Hong
H
Ma
Robert T
RT
Simpson
D018797:Cell Cycle Proteins / Q000378:metabolism / Q000502:physiology*; D002843:Chromatin / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism / Q000502:physiology*; D015825:Chromosomes, Fungal; D012441:Saccharomyces cerevisiae / Q000235:genetics; D000097722:Cohesins
D018797:Cell Cycle Proteins; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D000097722:Cohesins