Lysine methyltransferase SETD6 modifies histones on a glycine-lysine motif

Figures & data

Table 1. SETD6 consensus motif. Based on H2AZ methylation sites by SETD6, putative modification sites were identified in canonical histones H2A, H3, and H4.

Histone	consensus
	−3	−2	−1	0	+1	+2	+3
H2AZ	A	G	G	K^4	A	G	K
H2AZ	K	A	G	K^7	D	S	G
H2A	G	R	G	K^5	Q	G	G
H3	T	G	G	K^14	A	P	R
H4	G	R	G	K^5	G	G	K
H4	G	L	G	K^12	G	G	A
	•	•	*	*	•	:	•

Figure 1. SETD6 methylates histones. Recombinant KMTs were used to modify histones isolated from calf thymus in the presence of 3H-SAM. Samples were analysed by SDS-PAGE, then either stained with Coomassie (top panel) or transferred to PVDF membrane and autoradiographed (bottom panel).

Figure 2. SETD6 methylates GK motifs in canonical histones. (a) SETD6 was used to methylate the indicated recombinant histone tails. Samples were analysed by SDS-PAGE, then either stained with Coomassie (top panel) or transferred to PVDF membrane and autoradiographed (bottom panel). (b) KMT assays were performed with SETD6 on H4 peptides. (c) Sequence of H4 peptides used in panel B with each lysine numbered.

Table 2. Novel SETD6 putative substrates. The GKDS motif from H2AZ was used to identify novel substrates for SETD6.

Substrate	Site	Function
AHR	GK^438DS	Nuclear receptor
APAF1	GK^100DS	Apoptotis
CDKN2C	GK^160DS	CDK4 inhibitor p18^INK4C
GREB1	GK^278DS	Proliferation
KDM5A	GK^1504DS	Demethylase
MYST4	GK^393DS	Acetyltransferase
RBBP8	GK^426DS	pRB-binding protein
RUVBL1	GK^422DS	chromatin remodelling
SMYD4	GK^63DS	putative KMT
SUV420H1	GK^864DS	H4K20 KMT