Assembly dynamics of PML nuclear bodies in living cells

PMC Biophysics

Table 1 Exchange dynamics of PML isoforms at nuclear bodies

Protein	Cells	k_on(s^-1)	k_off(s^-1)	k_in(s^-1)	k_out(s^-1)	R.t. (s)	bnd_out	bnd_in
GFP-PML I	PML^+/+	0.0633	0.0051	0.0002	0.0004	300	0.65	0.35
GFP-PML I	PML^-/-	0.0015	0.0015	>0.0001	0.0049	648	1.00	0.00
GFP-PML II	PML^+/+	0.0856	0.0069	>0.0001	>0.0001	222	0.65	0.35
GFP-PML II^immob	PML^-/-	0.0000	n.d.	n.d.	n.d.	n.d.	n.d.	n.d.
GFP-PML II^mob	PML^-/-	0.0288	0.0031	>0.0001	>0.0001	659	0.49	0.51
GFP-PML III	PML^+/+	0.0692	0.0055	>0.0001	0.0007	274	0.67	0.33
GFP-PML III	PML^-/-	0.0023	0.0013	>0.0001	0.5592	744	1.00	0.00
GFP-PML IV	PML^+/+	0.0565	0.0034	>0.0001	>0.0001	337	0.87	0.13
GFP-PML IV^immob	PML^-/-	0.0040	0.0005	>0.0001	>0.0001	4777	0.38	0.62
GFP-PML IV^mob	PML^-/-	0.0571	0.0037	>0.0001	>0.0001	333	0.81	0.19
GFP-PML V	PML^+/+	0.0199	0.0024	>0.0001	>0.0001	957	0.43	0.57
GFP-PML V	PML^-/-	0.0188	0.0016	>0.0001	>0.0001	1010	0.61	0.39
GFP-PML VI	PML^+/+	0.0351	0.0021	>0.0001	>0.0001	541	0.90	0.10
GFP-PML VI	PML^-/-	0.0385	0.0036	0.0008	0.0010	493	0.56	0.44

The diffusion-binding model described in Figure 4A was used to extract kinetic data from the FRAP and FCS experiments for all PML isoforms in PML positive and PML negative cells. k_on: association rate at the surface of the nuclear body; k_off: dissociation rate at the surface of the nuclear body; k_in: penetration rate into the nuclear body; k_out: penetration rate out of the core of the nuclear body; R.t.: mean residence time at nuclear bodies; bnd_out: fraction of molecules residing at the surface of the nuclear body; bnd_in: fraction of molecules residing in the core of the nuclear body.