Systematic yeast synthetic lethal and synthetic dosage lethal screens identify genes required for chromosome segregation.
Accurate chromosome segregation requires the execution and coordination of many processes during mitosis, including DNA replication, sister chromatid cohesion, and attachment of chromosomes to spindle microtubules via the kinetochore complex. Additional pathways are likely involved because faithful chromosome segregation also requires proteins that are not physically associated with the chromosome. Using kinetochore mutants as a starting point, we have identified genes with roles in chromosome stability by performing genome-wide screens employing synthetic genetic array methodology. Two genetic approaches (a series of synthetic lethal and synthetic dosage lethal screens) isolated 211 nonessential deletion mutants that were unable to tolerate defects in kinetochore function. Although synthetic lethality and synthetic dosage lethality are thought to be based upon similar genetic principles, we found that the majority of interactions associated with these two screens were nonoverlapping. To functionally characterize genes isolated in our screens, a secondary screen was performed to assess defects in chromosome segregation. Genes identified in the secondary screen were enriched for genes with known roles in chromosome segregation. We also uncovered genes with diverse functions, such as RCS1, which encodes an iron transcription factor. RCS1 was one of a small group of genes identified in all three screens, and we used genetic and cell biological assays to confirm that it is required for chromosome stability. Our study shows that systematic genetic screens are a powerful means to discover roles for uncharacterized genes and genes with alternative functions in chromosome maintenance that may not be discovered by using proteomics approaches.
Measday
Vivien
V
Wine Research Centre and Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada V6T 1Z4.
Baetz
Kristin
K
Guzzo
Julie
J
Yuen
Karen
K
Kwok
Teresa
T
Sheikh
Bilal
B
Ding
Huiming
H
Ueta
Ryo
R
Hoac
Trinh
T
Cheng
Benjamin
B
Pot
Isabelle
I
Tong
Amy
A
Yamaguchi-Iwai
Yuko
Y
Boone
Charles
C
Hieter
Phil
P
Andrews
Brenda
B
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000378:metabolism; D043171:Chromosomal Instability / Q000235:genetics; D020090:Chromosome Segregation / Q000235:genetics*; D015825:Chromosomes, Fungal / Q000378:metabolism; D005800:Genes, Fungal; D005804:Genes, Lethal; D023281:Genomics / Q000379:methods; D018386:Kinetochores / Q000378:metabolism; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism
C090297:AFT1 protein, S cerevisiae; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; C064042:CBF1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors