Identification and characterization of a set of conserved and new regulators of cytoskeletal organization, cell morphology and migration

Bai, Siau Wei; Herrera-Abreu, Maria Teresa; Rohn, Jennifer L; Racine, Victor; Tajadura, Virginia; Suryavanshi, Narendra; Bechtel, Stephanie; Wiemann, Stefan; Baum, Buzz; Ridley, Anne J

doi:10.1186/1741-7007-9-54

Table 2 Cell morphology and cytoskeleton organisation in PMM-depleted PC3 cells.

From: Identification and characterization of a set of conserved and new regulators of cytoskeletal organization, cell morphology and migration

Group	Gene symbol	Actin	Microtubules	Cell shape	Spread area	Multi-nucleate
	control	stress fibres 4% strong patches 8% cortical actin 12% low 14%	processes 6%	elongated 10% rounded 4%	larger 4%	0%
1	ARC	stress fibres 70%	NP	elongated	larger	6%
	ZRANB1	stress fibres 90%	NP	elongated	larger	10%
2	C20orf142	cortical actin	NP	NP	larger 85%	NP
	CAMSAP1	cortical actin, spots	NP	NP	larger 85%	NP
	FMNL1	cortical actin 90%, rough edge	NP	NP	larger	NP
	FAM40A	cortical actin 90%, rough edge	NP	NP	NP	NP
	C21orf2	cortical actin 55%	NP	NP	NP	NP
3	FMNL2	cortical actin, spots	processes 60%	NP	larger	9%
	ZNF135	spots	processes 50%	NP	NP	NP
	ZMYM3	cortical actin, spots	processes 25%	NP	larger	NP
	PHIP	cortical actin	processes 35%	NP	NP	NP
4	FAM40B	cortical actin	processes	elongated 65%	NP	NP
	FNBP3	cortical actin, stress fibres	processes	elongated 65%	larger	4%
	FMNL3	NP	processes	elongated 80%	larger	NP
	Rac1	stress fibres	NP	elongated 65%	larger	NP
	LIMD1	NP	processes	elongated 50%	NP	NP
	SH3KBP1	NP	NP	elongated 90%	NP	NP
	RhoA	strong patches	NP	elongated 70%	NP	NP
	SH3D19	spots	NP	very elongated 90%	NP	NP
5	ZMYM4	spots	processes	very elongated 95%	NP	NP
	WAVE3	spots	processes	very elongated 90%	NP	NP
	LARP4	NP	processes	very elongated 45%	NP	NP
	DIAPH1	NP	processes	very elongated 90%	larger	NP
	RhoU	NP	NP	very elongated 80%	NP	NP
6	CAPZB	strong patches 90%, cortical actin	NP	NP	NP	NP
	CFL1	strong patches 80%	NP	NP	NP	31%
	PDZK8	strong patches 45%	NP	NP	NP	2%
7	ZMYM6	rough edge	NP	rounded 65%	NP	NP
	Cdc42	rough edge	processes	rounded 70%	NP	NP
	WTIP	NP	NP	rounded 95%	NP	NP
	BRWD1	NP	NP	rounded 80%	smaller	NP
	BRWD3	NP	NP	rounded 80%	smaller	NP
	HYPC	low 65%, smooth	NP	NP	NP	NP
	EPB41L4A	NP	NP	NP	NP	NP

Phenotypes were classified three days after transfection with siRNA pools targeting each of the 26 PMMs or 8 known regulators of the actin cytoskeleton (italics). Cells were fixed and stained for F-actin, microtubules, and DNA (as in Figure 1). Three to four different images from two different screens were analysed; total number of cells analysed/condition ≥ 68. The most prominent phenotype for each gene is highlighted in bold text and the percent of cells showing this phenotype is shown in brackets (see Figure 1, Supplementary. Figure 2 for examples of each phenotype). Definitions of actin phenotypes: stress fibres, multiple thick straight F-actin cables/cell; cortical actin, increased F-actin on regions of the plasma membrane; spots, small bright regions of F-actin; strong patches, large bright regions of F-actin; rough edge, free edges of cells are wavy or indented; smooth, F-actin levels similar throughout cytoplasm; low, lower F-actin levels than control cells. Microtubule phenotype: processes, long narrow membrane extensions containing microtubules. Definitions of cell shape: larger, spread area increased compared to control cells; elongated, cell body length ≥ 2 × width; very elongated, cell body length ≥ 3 × width; rounded, length ~ = width. NP, no phenotype.

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ISSN: 1741-7007

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