(a) reduced expression of Prmt1 in Prmt1+/- mice, compared to WT control mice. (b) Representative traces of seizure activities: short (upper) or long (middle and lower) duration of epileptiform …
Source data for Figure 1.
(a–b) a and b panels show representative trace in the whole-cell current-clamp recording from WT (a) and Prmt1+/- (b) mature dentate GCs in response to 1-s depolarizing current injection (200 pA), …
Source data for Figure 2.
(a–b) Representative current traces of voltage clamp recordings from WT (a; n = 22, seven mice) and Prmt1+/- (b; n = 12, four mice) mature dentate GCs in response to the voltage protocol depicted …
Source data for Figure 3.
Each connected line represents an individual neuron. Right panel gives the averaged changes. XE991 was less effective in mutants than in WT.
Source data for Figure 3—figure supplement 1.
(a–b) Representative current traces recorded from MTA- (a) or furamidine- (b) pretreated WT GCs using the voltage protocol depicted below in the absence (black) or presence (blue) of 10 μM XE991. (c)…
Source data for Figure 4.
(a–c) Immunoblotting analysis showing the physical association of KCNQ2 and Prmt1. HEK293T cells were transfected with expression vectors, as indicated. Whole-cell lysates were immunoprecipitated …
(a) 332RRNPAAGLIQSAWR345, (b) 334NPAAGLIQSAWR345, (c) 346FYATNLSR353 and (d) 435RSPSADQSLEDSPSK449. R* represents monomethylated arginine and R** represents dimethylated arginine.
The arginine residues critical for methylation in KCNQ2 are marked in red. KCNQ3 and KCNQ4 have 3 conserved arginine residues while KCNQ5 and KCNQ1 have two or one conserved arginine residues, …
In vitro methylation assays with myc-Prmt5 purified from HEK293T cells and GST-Prmt1 by using GST-KCNQ2 (aa 320–449) as a substrate and histone as a positive control. Coomassie blue stainings show …
(a) Representative current recordings from HEK293T cells expressing KCNQ2 with control shRNA vector (left) or a Prmt1 shRNA (Prmt1 sh) (right). Currents were elicited by voltage steps from -70 mV to …
Source data for Figure 6.
(a) Pooled data show 10 μM neomycin-induced rundown of WT (●) and R333K (○) KCNQ2 currents. KCNQ2 currents were normalized to KCNQ2 current at t = 0. (b) Dose-response curves for neomycin measured …
Source data for Figure 7.
Membrane surface biotinylation assays show that the ratio of channels on the membrane to those in the total lysates was not significantly different in control cells and Prmt1 knockdown cells (left). …
Tail current amplitudes were used to measure current inhibition by Dr-VSP activation and its recovery. (a–e) Time course of tail current amplitude in a cell transfected with Dr-VSP and WT KCNQ2 (a), …
Source data for Figure 7—figure supplement 2.
Changes in firing rate (a) and threshold current (b) in WT (+/+) neurons, mutants (+/-), and mutants loaded with 20μM diC8-PIP2 (+/-; PIP2) in response to 10 μM XE991. Each connected line represents …
Source data for Figure 7—figure supplement 3.
(a) representative trace in the whole-cell current-clamp recording from WT GCs with no PIP2 (left) or 20 μM diC8-PIP2 in the patch pipette (right) in response to 1-s depolarizing current injection …
Source data for Figure 7—figure supplement 4.
Spike trains were evoked by injecting 1-s depolarizing current pulses of different intensities (100–250 pA) into the cell before and after application of apamin (a–f) or paxilline (g–l) in control …
Source data for Figure 7—figure supplement 5.
Spike trains were evoked by injecting 1-s depolarizing current pulses of different intensities (100–250 pA) into the cell before (black) and after (red) application of H2O2. Left panels show …
Source data for Figure 8.
(a–g) Time course for the effect of 500 μM H2O2 on KCNQ2 currents. WT KCNQ2 (a-c), R333K (d), R345K (e), R353K (f) or R435K (g) were expressed in HEK293T cells. For experiment (b) & (c), cells were …
Source data for Figure 9.
Summarized data for Triple-Cys mutant of KCNQ2 currents activation induced by H2O2 or retigabine (right). (b) Treatment of MTA induces a reduction of KCNQ2(WT) and Triple-Cys mutant of KCNQ2 …
Source data for Figure 9—figure supplement 1.
Mono- and dimethylated peptides identified from KCNQ2.
Peptide* | Modification | Charge state |
---|---|---|
332RRNPAAGLIQSAWR345 | Mono-methylated R333 | +3 |
334NPAAGLIQSAWR345 | Mono-methylated R345 | +2 |
346FYATNLSR353 | Di-methylated R353 | +2 |
435RSPSADQSLEDSPSK449 | Di-methylated R435 | +3 |
*Methylated amino acid residues are highlighted in bold.