Identification of novel Wilms' tumor suppressor gene target genes implicated in kidney development.
The Wilms' tumor suppressor gene (WT1) encodes a zinc finger transcription factor that is vital during development of several organs including metanephric kidneys. Despite the critical regulatory role of WT1, the pathways and mechanisms by which WT1 orchestrates development remain elusive. To identify WT1 target genes, we performed a genome-wide expression profiling analysis in cells expressing inducible WT1. We identified a number of direct WT1 target genes, including the epidermal growth factor (EGF)-family ligands epiregulin and HB-EGF, the chemokine CX3CL1, and the transcription factors SLUG and JUNB. The target genes were validated using quantitative reverse transcriptase-polymerase chain reaction, small interfering RNA knockdowns, chromatin immunoprecipitation, and luciferase reporter analyses. Immunohistochemistry of fetal kidneys confirmed that a number of the WT1 target genes had overlapping expression patterns with the highly restricted spatiotemporal expression of WT1. Finally, using an in vitro embryonic kidney culture assay, we found that the addition of recombinant epiregulin, amphiregulin, CX3CL1, and interleukin-11 significantly enhanced ureteric bud branching morphogenesis. Our genome-wide screen implicates WT1 in the transcriptional regulation of the EGF-family of growth factors as well as the CX3CL1 chemokine during nephrogenesis.
Kim
Ho-Shik
HS
Genetics of Development and Disease Branch, Microarray Core Facility, NIDDK, National Institutes of Health, Bethesda, Maryland 20892, USA.
Kim
Myoung Shin
MS
Hancock
Anne L
AL
Harper
James C P
JC
Park
Jik Young
JY
Poy
George
G
Perantoni
Alan O
AO
Cam
Margaret
M
Malik
Karim
K
Lee
Sean Bong
SB
D066258:Amphiregulin; D000818:Animals; D045744:Cell Line, Tumor; D054360:Chemokine CXCL1; D019743:Chemokines, CXC / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D066255:EGF Family of Proteins; D004815:Epidermal Growth Factor / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D066260:Epiregulin; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology*; D016678:Genome / Q000502:physiology; D006023:Glycoproteins / Q000494:pharmacology; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology; D017370:Interleukin-11 / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000196:embryology*; D038081:Organogenesis / Q000187:drug effects / Q000502:physiology*; D011247:Pregnancy; D016755:Proto-Oncogene Proteins c-jun / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D025721:WT1 Proteins / Q000235:genetics / Q000378:metabolism*; D016335:Zinc Fingers / Q000235:genetics
C587848:AREG protein, human; D066258:Amphiregulin; C587850:Areg protein, rat; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; C513297:Cxcl1 protein, rat; D066255:EGF Family of Proteins; C587880:EREG protein, human; D066260:Epiregulin; C587882:Ereg protein, rat; D006023:Glycoproteins; C587883:HBEGF protein, human; C587885:Hbegf protein, rat; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D017370:Interleukin-11; D016755:Proto-Oncogene Proteins c-jun; C000604407:SNAI1 protein, human; C000604450:Snai2 protein, rat; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; D025721:WT1 Proteins; D004815:Epidermal Growth Factor