A systems approach reveals that the myogenesis genome network is regulated by the transcriptional repressor RP58.
We created a whole-mount in situ hybridization (WISH) database, termed EMBRYS, containing expression data of 1520 transcription factors and cofactors expressed in E9.5, E10.5, and E11.5 mouse embryos--a highly dynamic stage of skeletal myogenesis. This approach implicated 43 genes in regulation of embryonic myogenesis, including a transcriptional repressor, the zinc-finger protein RP58 (also known as Zfp238). Knockout and knockdown approaches confirmed an essential role for RP58 in skeletal myogenesis. Cell-based high-throughput transfection screening revealed that RP58 is a direct MyoD target. Microarray analysis identified two inhibitors of skeletal myogenesis, Id2 and Id3, as targets for RP58-mediated repression. Consistently, MyoD-dependent activation of the myogenic program is impaired in RP58 null fibroblasts and downregulation of Id2 and Id3 rescues MyoD's ability to promote myogenesis in these cells. Our combined, multi-system approach reveals a MyoD-activated regulatory loop relying on RP58-mediated repression of muscle regulatory factor (MRF) inhibitors.
Yokoyama
Shigetoshi
S
Department of Systems BioMedicine, National Research Institute for Child Health and Development, Setagaya, Tokyo 157-8535, Japan.
Ito
Yoshiaki
Y
Ueno-Kudoh
Hiroe
H
Shimizu
Hirohito
H
Uchibe
Kenta
K
Albini
Sonia
S
Mitsuoka
Kazuhiko
K
Miyaki
Shigeru
S
Kiso
Minako
M
Nagai
Akane
A
Hikata
Tomohiro
T
Osada
Tadahiro
T
Fukuda
Noritsugu
N
Yamashita
Satoshi
S
Harada
Daisuke
D
Mezzano
Valeria
V
Kasai
Masataka
M
Puri
Pier Lorenzo
PL
Hayashizaki
Yoshihide
Y
Okado
Haruo
H
Hashimoto
Megumi
M
Asahara
Hiroshi
H
D000818:Animals; D055785:Gene Knockdown Techniques; D055786:Gene Knockout Techniques; D053263:Gene Regulatory Networks; D051797:Inhibitor of Differentiation Protein 2 / Q000378:metabolism; D051796:Inhibitor of Differentiation Proteins / Q000378:metabolism; D051379:Mice; D024510:Muscle Development; D018482:Muscle, Skeletal / Q000196:embryology*; D018006:Myogenic Regulatory Factors / Q000235:genetics; D012097:Repressor Proteins / Q000378:metabolism*
C497452:Idb2 protein, mouse; D051797:Inhibitor of Differentiation Protein 2; D051796:Inhibitor of Differentiation Proteins; D018006:Myogenic Regulatory Factors; D012097:Repressor Proteins; C520669:Zbtb18 protein, mouse; C067679:Idb3 protein, mouse