A DNA fragment homologous to U6 small nuclear RNA was isolated from a human genomic library and sequenced. The immediate 5'-flanking region of the clone had significant homology with potential mouse gene, including "TATA box" at position 26-29 nucleotides upstream transcription start site. Although this sequence element is characteristic polymerase II promoters, gene also contained III "box A" intragenic control typical run five thymines 3' terminus (noncoding strand). accurately transcribed...

We examined the osteoblast/osteocyte expression and function of polycystin-1 (PC1), a transmembrane protein that is component polycystin-2 (PC2)-ciliary mechano-sensor complex in renal epithelial cells. found MC3T3-E1 osteoblasts MLO-Y4 osteocytes express transcripts for PC1, PC2, ciliary proteins Tg737 Kif3a. Immunohistochemical analysis detected cilia-like structures osteoblastic osteocyte-like cell lines as well primary from calvaria. Pkd1m1Bei mice have inactivating missense mutations...

Functional analysis of polycystin-1, the product gene most frequently mutated in autosomal dominant polycystic kidney disease, has revealed that this protein is involved regulation diverse signaling pathways such as activation transcription factor AP-1 and modulation Wnt signaling. However, initial steps cascades have remained unclear. We demonstrated previously C-terminal cytosolic tail polycystin-1 binds activates heterotrimeric G proteins <i>in vitro</i>. To test if can activate cellular...

Metanephric organ culture has been used to determine whether embryonic kidney tubules can be stimulated by cAMP form cysts. Under basal conditions, wild-type kidneys from day 13.5 15.5 mice grow in size and continue ureteric bud branching tubule formation over a 4- 5-d period. Treatment of these with 8-Br-cAMP or the agonist forskolin induced dilated within 1 h, which enlarged several days resulted dramatically expanded cyst-like structures proximal collecting duct origin. Tubule dilation...

Currently, there is little understanding of what factors regulate the development urine concentrating capability in normal or polycystic kidney. The present study examined developmental expression genes associated with concentration developing mice, including C57BL/6J-cpk/cpk mice autosomal recessive-infantile (AR) kidney disease (PKD). Concentration requires: 1) medullary collecting ducts (CD) located within a hypertonic interstitium, 2) CD cell functional arginine vasopressin V2 receptors...

U3 small nuclear RNA is hydrogen-bonded to high molecular weight nucleolar and can be isolated from greater than 60S pre-ribosomal ribonucleoprotein particles, suggesting that it involved in processing of ribosomal precursors (pre-rRNA) or ribosome biogenesis. Here we have used vivo psoralen cross-linking identify the region pre-rRNA interacting with RNA. Quantitative hybridization selection/depletion experiments clones rRNA-encoding DNA (rDNA) cross-linked showed all was associated a...

Polycystin-1 (Pc1) cleavage at the G protein-coupled receptor (GPCR) proteolytic site (GPS) is required for normal kidney morphology in humans and mice. We found a complex pattern of endogenous Pc1 forms by GPS cleavage. generates not only heterodimeric cleaved full-length (Pc1(cFL)) which N-terminal fragment (NTF) remains noncovalently associated with C-terminal (CTF) but also novel form (Pc1(deN)) NTF becomes detached from CTF. Uncleaved (Pc1(U)) resides primarily endoplasmic reticulum...

Oxidative stress has been implicated in the pathogenesis of both acquired and hereditary polycystic kidney disease. Mechanisms oxidant injury C57BL/6J-cpk mice Han:SPRD-Cy rats with rapidly or slowly progressive disease were explored. Expression heme oxygenase-1 mRNA, an inducible marker oxidative stress, was shown to be increased cystic kidneys a pattern that reflected severity. By contrast, there decrease mRNA expression antioxidant enzymes extracellular glutathione peroxidase, superoxide...

Polycystin-1, the protein product of polycystic kidney disease-1 (PKD1) gene, was originally predicted to be an integral membrane glycoprotein with 11 transmembrane (TM) domains (TM 1-11). Subsequent comparative sequence analyses led a revision original model, which retained overall topology and TM segments I-XI) but dropped 3 introduced new domains. The membrane-spanning potential orientation each proposed following extracellular REJ domain I-XI 11) have now been tested. Using series...

A mouse kidney cDNA isolated by differential screening was found to be highly homologous rat, human, and bovine plasma glutathione peroxidase (GPx) sequences. Analysis of the full-length coding region sequence demonstrated an in-frame selenocysteine-encoding opal codon putative signal sequence, suggesting that represents homolog GPx. The level expression GPx in various tissues during development investigated Northern blot analysis. Plasma mRNA observed very abundant compared with placenta,...

In polycystic kidney disease (PKD), renal parenchyma is destroyed by cysts, hypothesized to obstruct nephrons. A signature of unilateral ureteral obstruction, proximal tubular atrophy leads formation atubular glomeruli. To determine whether this process occurs in PKD, kidneys from pcy mice (moderately progressive PKD), cpk (rapidly and human autosomal dominant PKD were examined early late stages. Integrity the glomerulotubular junction mass determined sections stained with Lotus...

Polycystic kidney disease (PKD) is a genetic disorder characterized by fluid-filled cysts in the and liver that ultimately leads to end-stage renal disease. Currently there no globally approved therapy for PKD. The Notch signaling pathway regulates cellular processes such as proliferation de-differentiation, which are hallmarks of Thus we hypothesized plays critical role Evaluation protein expression components kidneys Autosomal Recessive PKD (ARPKD) Dominant (ADPKD) mouse models ADPKD...

Autosomal dominant polycystic kidney disease (ADPKD) is characterized by the growth of renal cysts that ultimately destroy function. Mutations in PKD1 and PKD2 genes cause ADPKD. Their protein products, polycystin-1 (PC1) polycystin-2 (PC2) have been proposed to form a calcium-permeable receptor-channel complex; however mechanisms which they function are almost completely unknown. Most mutations truncating loss-of-function or affect biogenesis, trafficking stability reveal very little about...

Polycystic kidney disease (PKD) results from loss-of-function mutations in the PKD1 gene. There are also reports showing abnormally high levels of expression cystic epithelial cells. At present, nothing is known about molecular mechanisms regulating normal thePKD1 gene or whether transcriptional disregulation has a role cyst formation. We have analyzed 3.3-kb 5′-proximal portion human PKD1gene. Sequence analysis revealed presence consensus sequences for numerous transactivating factors,...

Significance Mutations of polycystin-1 (PC1) are the major cause (85% cases) autosomal dominant polycystic kidney disease (ADPKD), which is fourth leading failure. PC1 thought to function as an atypical G protein-coupled receptor, yet mechanism by regulates G-protein signaling remains poorly understood. A significant portion ADPKD mutations encode a protein with defects in maturation or reduced that may be amenable functional rescue. In this work, we have combined complementary biochemical...

Polycystic kidney diseases (PKD) are inherited as autosomal dominant (ADPKD) or recessive (ARPKD) traits and characterized by progressive enlargement of renal cysts. Aberrant cell proliferation is a key feature in the progression PKD. Cux1 homeobox gene that related to Drosophila cut murine homolog human CDP (CCAAT Displacement Protein). represses cyclin kinase inhibitors p21 p27, transgenic mice ectopically expressing develop hyperplasia. However, do not Here, we show 246 amino acid...