The histone fold domain of Cse4 is sufficient for CEN targeting and propagation of active centromeres in budding yeast.
Centromere-specific H3-like proteins (CenH3s) are conserved across the eukaryotic kingdom and are required for packaging centromere DNA into a specialized chromatin structure required for kinetochore assembly. Cse4 is the CenH3 protein of the budding yeast Saccharomyces cerevisiae. Like all CenH3 proteins, Cse4 consists of a conserved histone fold domain (HFD) and a divergent N terminus (NT). The Cse4 NT contains an essential domain designated END (for essential N-terminal domain); deletion of END is lethal. To investigate the role of the Cse4 NT in centromere targeting, a series of deletion alleles (cse4DeltaNT) were analyzed. No part of the Cse4 NT was required to target mutant proteins to centromere DNA in the presence of functional Cse4. A Cse4 degron strain was used to examine targeting of a Cse4DeltaNT protein in the absence of wild-type Cse4. The END was not required for centromere targeting under these conditions, confirming that the HFD confers specificity of Cse4 centromere targeting. Surprisingly, overexpression of the HFD bypassed the requirement for the END altogether, and viable S. cerevisiae strains in which the cells express only the Cse4 HFD and six adjacent N-terminal amino acids (Cse4Delta129) were constructed. Despite the complete absence of the NT, mitotic chromosome loss in the cse4Delta129 strain increased only 6-fold compared to a 15-fold increase in strains overexpressing wild-type Cse4. Thus, when overexpressed, the Cse4 HFD is sufficient for centromere function in S. cerevisiae, and no posttranslational modification or interaction of the NT with other kinetochore component(s) is essential for accurate chromosome segregation in budding yeast.
Morey
Lisa
L
Department of Molecular Genetics and Microbiology, University of Massachusetts Medical School, 55 Lake Ave. North, Worcester, MA 01655, USA.
Barnes
Kelly
K
Chen
Yinhuai
Y
Fitzgerald-Hayes
Molly
M
Baker
Richard E
RE
D000483:Alleles; D015152:Blotting, Northern; D015139:Blotting, Southern; D002503:Centromere / Q000737:chemistry / Q000648:ultrastructure; D002843:Chromatin / Q000737:chemistry; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry*; D005656:Fungal Proteins / Q000737:chemistry*; D017353:Gene Deletion; D005816:Genetic Complementation Test; D005838:Genotype; D005947:Glucose / Q000378:metabolism; D006657:Histones / Q000737:chemistry*; D018386:Kinetochores / Q000737:chemistry / Q000378:metabolism; D008954:Models, Biological; D008957:Models, Genetic; D009154:Mutation; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013997:Time Factors
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; C467729:Cse4 protein, Candida albicans; D005656:Fungal Proteins; D006657:Histones; D005947:Glucose