A unique stylopod patterning mechanism by Shox2-controlled osteogenesis.
Vertebrate appendage patterning is programmed by Hox-TALE factor-bound regulatory elements. However, it remains unclear which cell lineages are commissioned by Hox-TALE factors to generate regional specific patterns and whether other Hox-TALE co-factors exist. In this study, we investigated the transcriptional mechanisms controlled by the Shox2 transcriptional regulator in limb patterning. Harnessing an osteogenic lineage-specific Shox2 inactivation approach we show that despite widespread Shox2 expression in multiple cell lineages, lack of the stylopod observed upon Shox2 deficiency is a specific result of Shox2 loss of function in the osteogenic lineage. ChIP-Seq revealed robust interaction of Shox2 with cis-regulatory enhancers clustering around skeletogenic genes that are also bound by Hox-TALE factors, supporting a lineage autonomous function of Shox2 in osteogenic lineage fate determination and skeleton patterning. Pbx ChIP-Seq further allowed the genome-wide identification of cis-regulatory modules exhibiting co-occupancy of Pbx, Meis and Shox2 transcriptional regulators. Integrative analysis of ChIP-Seq and RNA-Seq data and transgenic enhancer assays indicate that Shox2 patterns the stylopod as a repressor via interaction with enhancers active in the proximal limb mesenchyme and antagonizes the repressive function of TALE factors in osteogenesis.
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA.
Wenduo
W
Ye
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA Southern Center for Biomedical Research and Fujian Key Laboratory of Developmental and Neural Biology, College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, People's Republic of China.
Yingnan
Y
Song
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA Southern Center for Biomedical Research and Fujian Key Laboratory of Developmental and Neural Biology, College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, People's Republic of China.
Zhen
Z
Huang
Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Marco
M
Osterwalder
Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
Anja
A
Ljubojevic
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA State Key Laboratory of Oral Disease, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan 610041, People's Republic of China.
Jue
J
Xu
Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
Brent
B
Bobick
Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
Samuel
S
Abassah-Oppong
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA Southern Center for Biomedical Research and Fujian Key Laboratory of Developmental and Neural Biology, College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, People's Republic of China.
Ningsheng
N
Ruan
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA.
Ross
R
Shamby
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA.
Diankun
D
Yu
Department of Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Lu
L
Zhang
Department of Developmental and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Chen-Leng
CL
Cai
Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA U.S. Department of Energy Joint Genome Institute, Walnut Creek, CA 94598, USA School of Natural Sciences, University of California at Merced, Merced, CA 95343, USA.
Axel
A
Visel
Southern Center for Biomedical Research and Fujian Key Laboratory of Developmental and Neural Biology, College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, People's Republic of China.
Yanding
Y
Zhang
Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada T2N 1N4.
John
J
Cobb
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA Southern Center for Biomedical Research and Fujian Key Laboratory of Developmental and Neural Biology, College of Life Sciences, Fujian Normal University, Fuzhou, Fujian 350108, People's Republic of China ychen@tulane.edu.
YiPing
0000-0002-8628-7713
Y
Chen
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D019521:Body Patterning / Q000235:genetics; D019070:Cell Lineage; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005121:Extremities / Q000196:embryology*; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D016678:Genome; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D059372:Nucleotide Motifs / Q000235:genetics; D010012:Osteogenesis / Q000235:genetics; D011485:Protein Binding
D050659:Core Binding Factor Alpha 1 Subunit; D018398:Homeodomain Proteins; C484663:Shox2 protein, mouse
Limb; Patterning; Shox2; Skeleton; Stylopod