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Chlamydomonas

Cyclin B-GFP in dividing cells
EB1-NeonGreen in dividing cells
EB1-NeonGreen in cells blocked by apc-mutation

 

Yeast

Driving synchronized cell cycles with periodic pulses of the anaphase inducer Cdc20.
The essential gene CDC20 is placed under control of the galactose inducible GAL1 promoter. The microfluidics flow-cell device of Charvin et al.( PLoS ONE. 2008 Jan 23;3(1):e1468) was used to provide periodic pulses of galactose at a period sufficient for each blocked cell to complete its cell cycle and reaccumulate at a new cdc20-deficient block. Green: tubulin (TUB1-YFP) marking mitotic spindle; yellow: CDC10-YFP: marker of bud neck, appears at budding and disappears at cytokinesis. Bottom: bars indicate periods of induction with galactose.
Periodic Start driven in a flow cell
Cells deleted for all G1 cyclins CLN1, CLN2 and CLN3 but provided with a methionine-regulatable MET3-CLN2 gene were periodically pulsed with methionine-free medium in the microfluidics flow-cell device of Charvin et al.( PLoS ONE. 2008 Jan 23;3(1):e1468). Green: Whi5-GFP; G1/S transcriptional repressor. Yellow: CDC10-YFP: marker of bud neck, appears at budding and disappears at cytokinesis. Bottom: bars indicate periods of induction with -Met.
Wild-type budding yeast cell cycle
Septin (CDC10-YFP) marks bud neck and disappears at cytokinesis; tubulin (TUB1-GFP) marks spindle; elongates at anaphase and disassembles at telophase.
Daughter-specific gene expression
The CTS1 promoter driving YFP produces gene expression, and resulting yellow fluorescence, in daughter cells exclusively, due to the asymmetric transcription factor Ace2.
Daughter-specific transcription factor Ace2
The Ace2 transcription factor is asymmetrically inherited, with significance residence only in the daughter nucleus.
Measuring size and the cell cycle
The constitutive ACT1 promoter drives red fluorescent protein DsRed; Myo1-GFP marks the bud neck in green. The Myo1 ring disappears at cytokinesis. The quantity of red fluorescence per cell was used as a surrogate for total cell protein content. Reference: Di Talia et al, Nature, 2007.
G1/S-specific expression from the CLN2 promoter
The G1/S-regulated promoter for the CLN2 G1 cyclin drives unstable GFP. Gene expression (pulse of green fluorescence) is detectable in every cell cycle, starting around the time of budding.
Nuclear localization of Whi5-GFP, a repressor of G1/S-specific gene
Whi5-GFP yields brief periods of nuclear green fluorescence immediately following cell division. Note that nuclear residence of Whi5 is longer in daughters than in mothers.

 





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Laboratory of Cell Cycle Genetics
The Rockefeller University
1230 York Avenue
New York, NY 10065

Fred Cross
Professor