Fcp1 phosphatase controls Greatwall kinase to promote PP2A-B55 activation and mitotic progression
- CEINGE Biotecnologie Avanzate, Italy
- University of Naples Federico II, Italy
- Consiglio Nazionale delle Ricerche, Italy
Figures
Figure 1 with 4 supplements
Fcp1 affects PP2A-B55-dependent dephosphorylations at mitosis exit.
(A, B) Control (Cont.) or Fcp1-depleted (Fcp1) by siRNAs, as well as Fcp1-depleted complemented with wild type Fcp1 (Fcp1WT), HeLa cells were arrested at prometaphase. (A) Cells were collected and …
Figure 1—figure supplement 1
Fcp1-dependency of mitotic exit dephosphorylations.
Control (Cont.) or Fcp1-depleted (Fcp1), as well as Fcp1-depleted complemented with wild type Fcp1 (Fcp1WT), HeLa cells were arrested at prometaphase. Lysates from cells released from prometaphase …
Figure 1—figure supplement 2
Okadaic acid-sensitive mitotic exit dephosphorylations.
Prometaphase-arrested HeLa cells were collected and, immediately after taking the time 0 sample, split into two portions, one portion received vehicle (Cont.) and the other 500 nM okadaic acid (OA); …
Figure 1—figure supplement 3
PRC1 localization in Fcp1-depleted cells.
(A) Asynchronously growing control (Cont. siRNAs) or Fcp1-siRNAs-treated (Fcp1 siRNAs) HeLa cells were fixed and stained for PRC1 (red), α-tubulin (green) and DNA (blue). (B) Control (Cont. siRNAs) …
Figure 1—figure supplement 4
Mitotic phenotypes in Fcp1-depleted cells.
Asynchronously growing control-siRNAs- (Cont. siRNAs) or Fcp1-siRNAs-treated (Fcp1 siRNAs) HeLa cells were taken at the indicated time points (hours, hrs) from the siRNAs treatment. Upper left …
Figure 2 with 2 supplements
Fcp1-dependency of Gwl dephosphorylation at S90 and S453 during mitosis exit.
(A) Ensa IP from prometaphase-arrested HeLa cells, previously transfected with a Flag-hEnsa expression vector, was divided into four sets and incubated as substrate in phosphatase reactions with …
Figure 2—figure supplement 1
Cdk1-dependent phosphorylations of Gwl.
(A) Mock- (Mk), V5-GwlWT-, V5-GwlS90A- and V5-GwlS453A-transfected HeLa cells were arrested at prometaphase, lysed and processed for V5 IP. IPs were separated in parallel SDS/PAGE and probed for …
Figure 2—figure supplement 2
Effect of prolonged Cdk1 inhibition on Gwl dephosphorylation in Fcp1-depleted cells.
Control (Cont.) or Fcp1-depleted (Fcp1) HeLa cells were arrested at prometaphase, split into two samples, one sample receiving vehicle (-), the other RO3306 (+), and further incubated for 30 min. …
Figure 3 with 1 supplement
Fcp1 binds and dephosphorylates Gwl during mitosis exit.
(A) Prometaphase-arrested HeLa cells were released into fresh medium and sampled at indicated time points of incubation. Upper panels, cell lysates were processed for mock (Mk) or Gwl IP that were …
Figure 3—figure supplement 1
Fcp1-Gwl interaction in hTERT-RPE1 cells.
hTERT-RPE1 cells were arrested at prometaphase, collected, released from arrest and sampled at the indicated time points during further incubation in fresh medium. Mock (Mk) or Gwl …
Figure 4 with 2 supplements
Fcp1-dependent dephosphorylation reduces Gwl kinase activity towards Ensa.
(A) Gwl IP from prometaphase-arrested HeLa cell lysates were divided into three sets and incubated for phosphatase reactions with either just buffer (Cont.), Fcp1WT or Fcp1CD proteins (Mk IP; 1/3 …
Figure 4—figure supplement 1
Fcp1 affects Ensa/ARPP19 kinase ability of Gwl.
(A) V5 IP from V5-GwlWT-transfected, prometaphase-arrested, HeLa cells was divided into three sets and incubated for phosphatase reactions with buffer (Cont.), Fcp1WT or Fcp1CD proteins (Mk IP; 1/3 …
Figure 4—figure supplement 2
Effects of prolonged Cdk1 inhibition on Gwl activity in Fcp1-depleted cells.
Gwl IPs from cell lysates of prometaphase-arrested Fcp1 siRNAs-treated (Fcp1) HeLa cells treated – or + RO3306 for 30 min, the same cell lysates of the experiment described in Figure 2—figure …
