Identification of NR5A1 (SF-1/AD4BP) gene expression modulators by large-scale gain and loss of function studies.
Nuclear receptor subfamily 5, group A, member 1 (NR5A1 previously known as SF-1/AD4BP) is a transcription factor involved in the development of adrenal/gonadal tissues and steroidogenic lineage cell differentiation in adult somatic stem cells. To understand the cellular signaling network that regulates NR5A1 gene expression, loss of function screening with an siRNA kinome library, and gain of function screening with an addressable full-length cDNA library representing one quarter of the human genome was carried out. The NR5A1 gene expression was activated in mesenchymal stem cells by siRNA directed against protein kinase C (PKC)-delta, erb-B3, RhoGAP (ARHGAP26), and hexokinase 2, none of which were previously known to be involved in the NR5A1 gene expression. Among these, we identified crosstalk between erb-B3 and PKC-delta signaling cascades. In addition, the gain of function studies indicated that sex-determining region Y (SRY)-box 15 (SOX15), TEA domain family member 4, KIAA1257 (a gene of unknown function), ADAM metallopeptidase with thrombospondin type 1 motif 6, Josephin domain containing 1, centromere protein, TATA box-binding protein-associated factor 5-like RNA polymerase, and inducible T-cell co-stimulator activate NR5A1 gene expression. These results provide new insights into the molecular mechanisms of NR5A1 gene expression.
Sakai
Noriko
N
Department of Stem Cell Based Drug Discovery, Research Center Kobe, Bayer Yakuhin Ltd, Kobe, Hyogo 650-0047, Japan.
Terami
Hiromi
H
Suzuki
Shinobu
S
Haga
Megumi
M
Nomoto
Ken
K
Tsuchida
Nobuko
N
Morohashi
Ken-ichirou
K
Saito
Naoaki
N
Asada
Maki
M
Hashimoto
Megumi
M
Harada
Daisuke
D
Asahara
Hiroshi
H
Ishikawa
Tetsuya
T
Shimada
Fumiki
F
Sakurada
Kazuhiro
K
D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D000242:Cyclic AMP / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D051745:Protein Kinase C-delta / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D020893:Receptor, ErbB-3 / Q000235:genetics / Q000378:metabolism; D055747:SOX Transcription Factors / Q000235:genetics / Q000378:metabolism; D054339:Steroidogenic Factor 1 / Q000235:genetics* / Q000378:metabolism*; D000091262:TEA Domain Transcription Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D002868:Chromosomal Proteins, Non-Histone; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D020690:GTPase-Activating Proteins; D009124:Muscle Proteins; D034741:RNA, Small Interfering; D055747:SOX Transcription Factors; C102810:SOX15 protein, human; D054339:Steroidogenic Factor 1; D000091262:TEA Domain Transcription Factors; C486309:TEAD4 protein, human; D014157:Transcription Factors; C085379:rho GTPase-activating protein; D000242:Cyclic AMP; D006593:Hexokinase; D020893:Receptor, ErbB-3; D051745:Protein Kinase C-delta; D051722:ADAM Proteins