Transcription factor HNF1beta and novel partners affect nephrogenesis.
Heterozygous mutations of the tissue-specific transcription factor hepatocyte nuclear factor (HNF)1beta, cause maturity onset diabetes of the young (MODY5) and kidney anomalies including agenesis, hypoplasia, dysplasia and cysts. Because of these renal anomalies, HNF1beta is classified as a CAKUT (congenital anomalies of the kidney and urinary tract) gene. We searched for human fetal kidney proteins interacting with the N-terminal region of HNF1beta using a bacterial two-hybrid system and identified five novel proteins along with the known partner DCoH. The interactions were confirmed for four of these proteins by GST pull-down assays. Overexpression of two proteins, E4F1 and ZFP36L1, in Xenopus embryos interfered with pronephros formation. Further, in situ hybridization showed overlapping expression of HNF1beta, E4F1 and ZFP36L1 in the developing pronephros. HNF1beta is present largely in the nucleus where it colocalized with E4F1. However, ZFP36L1 was located predominantly in the cytoplasm. A nuclear function for ZFP36L1 was shown as it was able to reduce HNF1beta transactivation in a luciferase reporter system. Our studies show novel proteins may cooperate with HNF1beta in human metanephric development and propose that E4F1 and ZFP36L1 are CAKUT genes. We searched for mutations in the open reading frame of the ZFP36L1 gene in 58 patients with renal anomalies but found none.
Dudziak
Karin
K
Institut für Zellbiologie Tumorforschung, Universitätsklinikum Essen, Universität Duisburg-Essen, Essen, Germany.
Mottalebi
Nima
N
Senkel
Sabine
S
Edghill
Emma L
EL
Rosengarten
Stefan
S
Roose
Magdalena
M
Bingham
Coralie
C
Ellard
Sian
S
Ryffel
Gerhart U
GU
D000818:Animals; D051817:Butyrate Response Factor 1 / Q000235:genetics / Q000378:metabolism*; D004252:DNA Mutational Analysis; D004625:Embryo, Nonmammalian / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D051539:Hepatocyte Nuclear Factor 1-beta / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000002:abnormalities / Q000196:embryology* / Q000378:metabolism; D038081:Organogenesis / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases; D014981:Xenopus
D051817:Butyrate Response Factor 1; C496495:HNF1B protein, human; D012097:Repressor Proteins; C102034:ZFP36L1 protein, human; D051539:Hepatocyte Nuclear Factor 1-beta; C513814:E4F1 protein, human; D044767:Ubiquitin-Protein Ligases