Description

This 547bp sequence, located in an intronic region of KH.L128.2, drives reporter gene expression in mesenchyme and central nervous system. It contains 1 Myb#2, 6 Brachyury, 2 Fox, 3 E-box, 1 Myb#3 and 1 Sp1/Klf putative binding sites. Compared to the notochord-active WT enhancer, it lacks the 4th Brachyury binding site.

Hierarchy of Regulatory Regions

Modification of Cirobu.REG.KH2012.L128:214487-215033|Furin,Nec1/2 by substitution

Regulated gene(s)

KH2012:KH.L128.2 (FURIN; PCSK1; PCSK2)

Construct(s) made to test this region

CONS00001394: p.Cirobu.REG.KH2012.L128:214487-215033_Bra4Mut2>LacZ

Overview of functional TF binding sites in this region

214,487215,033

	Motif Name	Binding Factor(s)	Sequence	Position in Region	Comment
1	Helix Turn Helix / Tryptophane clusters	n/a	ccgttg	[214,502 / 214,508]	This Myb#3 binding site is located on the notochord-active intronic enhancer of KH.L128.2.
2	T-Box	n/a	tagcac	[214,578 / 214,584]	This is the second of the 8 Brachyury binding sites located on the notochord-active intronic enhancer of KH.L128.2.
3	bHLH	n/a	cacatg	[214,581 / 214,587]	This is the second of the 4 E-box binding sites located on the notochord-active intronic enhancer of KH.L128.2.
4	bHLH	n/a	catttg	[214,593 / 214,599]	This is the third of the 4 E-box binding sites located on the notochord-active intronic enhancer of KH.L128.2.
5	ZnF_(C2H2)	n/a	ccacc	[214,616 / 214,621]	This Sp1/Klf binding site is located on the notochord-active intronic enhancer of KH.L128.2.
6	T-Box	n/a	tttcac	[214,633 / 214,639]	This is the third of the 8 Brachyury binding sites located on the notochord-active intronic enhancer of KH.L128.2.
7	bHLH	n/a	cagatg	[214,647 / 214,653]	This is the fourth of the 4 E-box binding sites located on the notochord-active intronic enhancer of KH.L128.2.
8	T-Box	n/a	ttacac	[214,697 / 214,703]	This is the fifth of the 8 Brachyury binding sites located on the notochord-active intronic enhancer of KH.L128.2.
9	Fox	n/a	tatttgc	[214,735 / 214,742]	This is the second of the 2 Fox binding sites located on the notochord-active intronic enhancer of KH.L128.2.
10	T-Box	n/a	gtggaa	[214,782 / 214,788]	This is the 6th of the 8 Brachyury binding sites located on the notochord-active intronic enhancer of KH.L128.2.
11	T-Box	n/a	gtgaaa	[214,801 / 214,807]	This is the 7th of the 8 Brachyury binding sites located on the notochord-active intronic enhancer of KH.L128.2.
12	Fox	n/a	ataaaca	[214,864 / 214,871]	This is the third of the 3 Fox binding sites located on the notochord-active intronic enhancer of KH.L128.2. Its mutation does not affect enhancer activity in the notochord, but it increases activity in the mesenchyme.
13	Helix Turn Helix / Tryptophane clusters	n/a	attgaa	[214,917 / 214,923]	This is the second of the 2 Myb#2 binding sites located on the notochord-active intronic enhancer of KH.L128.2.
14	T-Box	n/a	gtgcca	[214,965 / 214,971]	This is the 8th of the 8 Brachyury binding sites located on the notochord-active intronic enhancer of KH.L128.2. Its mutation decreases but does not inhibit enhancer activity in the notochord.

Sequence

Aniseed Coordinates: [214,487 / 215,033] on scaffold KhL128

This sequence was retrieved using ANISEED Blast tool, thanks to the information provided in FigS3.c and TableS5 of Jose-Edwards et al. 2015. The 4th Brachyrury binding site is mutated. GTGTCA replaced by AGATCA.

Reference(s)

José-Edwards et al. (2015). Brachyury, Foxa2 and the cis-Regulatory Origins of the Notochord. PLoS Genet. 11(12):e1005730