A new enrichment approach identifies genes that alter cell cycle progression in Saccharomyces cerevisiae.
Mechanisms that coordinate cell growth with division are thought to determine the timing of initiation of cell division and to limit overall cell proliferation. To identify genes involved in this process in Saccharomyces cerevisiae, we describe a method that does not rely on cell size alterations or resistance to pheromone. Instead, our approach was based on the cell surface deposition of the Flo1p protein in cells having passed START. We found that over-expression of HXT11 (which encodes a plasma membrane transporter), PPE1 (coding for a protein methyl esterase), or SIK1 (which encodes a protein involved in rRNA processing) shortened the duration of the G1 phase of the cell cycle, prior to the initiation of DNA replication. In addition, we found that, although SIK1 was not part of a mitotic checkpoint, SIK1 over-expression caused spindle orientation defects and sensitized G2/M checkpoint mutant cells. Thus, unlike HXT11 and PPE1, SIK1 over-expression is also associated with mitotic functions. Overall, we used a novel enrichment approach and identified genes that were not previously associated with cell cycle progression. This approach can be extended to other organisms.
Bogomolnaya
Lydia M
LM
Department of Biochemistry and Biophysics, Texas A&M University, 2128 TAMU, College Station, TX 77843, USA.
Pathak
Ritu
R
Cham
Roxana
R
Guo
Jinbai
J
Surovtseva
Yulia V
YV
Jaeckel
Lane
L
Polymenis
Michael
M
D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D015966:Gene Expression Regulation, Fungal / Q000235:genetics / Q000502:physiology; D005800:Genes, Fungal; D015698:Genomic Library; D037241:Mannose-Binding Lectins; D008938:Mitosis / Q000235:genetics* / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism
D018797:Cell Cycle Proteins; C087578:FLO1 protein, S cerevisiae; D037241:Mannose-Binding Lectins; D029701:Saccharomyces cerevisiae Proteins