Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.26.9 (ribonuclease)
 6,589 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have isolated the Penicillium chrysogenum nre gene which is homologous to the major nitrogen regulatory genes areA from Aspergillus nidulans and nit-2 from Neurospora crassa. Overall, nre shows 60% identity to areA and 30% identity to nit-2 at the amino-acid level. The gene encodes a protein of 835 amino-acid residues and contains a single Cys2/Cys2-type zinc finger with an adjacent basic region and a putative acidic activation region. In the DNA-binding domain, 98% of the amino-acid residues are identical in nre, areA and nit-2. The nre gene has been shown to be functional in N. crassa by heterologous complementation of a nit-2 mutant. Growth tests indicated that transformants could utilize nitrate, amino-acids, purines and amides as sole nitrogen sources. Nitrate reductase activity assays performed with transformants demonstrated that nitrogen control was completely normal. Complementation of N. crassa nit-2 mutants with 5'-deletion clones of nre suggests the possible presence of an internal promoter within the coding region. Northern analysis and ribonuclease protection assays of total cellular RNA indicated that nre encodes a 3.2-kb transcript which is reduced in content under conditions of nitrogen repression.
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PMID:Molecular cloning and analysis of nre, the major nitrogen regulatory gene of Penicillium chrysogenum. 778 18

Hepatocyte nuclear factor 6 (HNF-6) is the prototype of a family of tissue-specific transcription factors characterized by a bipartite DNA-binding domain consisting of a single cut domain and a novel type of homeodomain. We have previously cloned rat cDNA species coding for two isoforms, HNF-6alpha (465 residues) and beta (491 residues), which differ only by the length of the spacer between the two DNA-binding domains. We have now localized the rat Hnf6 gene to chromosome 8q24-q31 by Southern blotting of DNA from somatic cell hybrids and by fluorescence in situ hybridization. Cloning and sequencing of the rat gene showed that the two HNF-6 isoforms are generated by alternative splicing of three exons that are more than 10 kb apart from each other. Exon 1 codes for the N-terminal part and the cut domain, exon 2 codes for the 26 HNF-6beta-specific amino acids, and exon 3 codes for the homeodomain and the C-terminal amino acids. The transcription initiation site was mapped by ribonuclease protection and 5' rapid amplification of cDNA ends. Transfection experiments showed that promoter activity was contained within 0.75 kb upstream of the transcription initiation site. This activity was detected by the transfection of liver-derived HepG2 cells, but not of Rat-1 fibroblasts, suggesting that the promoter is sufficient to confer liver-specific expression.
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PMID:Hepatocyte nuclear factor 6: organization and chromosomal assignment of the rat gene and characterization of its promoter. 972 63

The expression time course of estrogen receptor alpha (ER alpha) was analyzed by RT-PCR in fetal and newborn rat pituitaries. In addition to the classical ER alpha messenger RNA (mRNA), three shorter transcripts were detected and subsequently cloned. Sequence analysis showed that they corresponded to ER alpha mRNAs lacking exon 3 (which encodes a zinc finger in the DNA-binding domain), exon 4 (which encodes the nuclear localization signal and part of the steroid-binding domain), or both exons 3 and 4. As analyzed by RT-PCR and ribonuclease protection assay, the respective expression levels of the different transcripts varied dramatically during pituitary development; short forms appeared 4 days before full-length ER alpha mRNA. On Western blots from rat pituitaries of different ages, an ER alpha-specific antiserum labeled four protein bands of the expected molecular weights, revealing that all four ER alpha mRNAs are translated in vivo. Immunocytochemistry, using the same antiserum, showed the ER alpha to be present first in the cytosol of intermediate lobe cells (around embryonic day 16). Only 5 days later, nuclear staining became detectable in the anterior lobe. We argue that the observed cytosolic staining will be essentially due to short ER alpha isoforms, which are indeed more abundantly expressed in the intermediate lobe. These data suggest that during pituitary development, the activity of the ER alpha might be specifically regulated by differential splicing of its primary transcript, resulting in a differential subcellular localization of the isoforms.
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PMID:Stage- and region-specific expression of estrogen receptor alpha isoforms during ontogeny of the pituitary gland. 1034 69

A DNA sequence corresponding to most of the DNA-binding domain of a Lucilia cuprina ultraspiracle protein (LcUSP) was amplified by PCR from genomic DNA and cloned. This cloned fragment was used to screen a L. cuprina cDNA library and to isolate a full-length LcUSP encoding sequence within a 3800-bp cDNA clone. The conceptually translated amino acid sequence of this open reading frame (467 amino acids) was used in alignment comparisons and phylogenetic analyses to reveal that LcUSP most closely resembles DmUSP relative to other known insect nuclear hormone receptors. An antisense RNA probe specific for the 5' end of Lcusp was used in ribonuclease protection assays to detect significant levels of Lcusp RNA throughout L. cuprina development. Highest levels were detected in embryos, late third instar larvae, pupae and adult females. This pattern parallels the pattern of expression observed for Dmusp RNAs during Drosophila melanogaster development. Finally, the LcUSP sequence was engineered for expression in mammalian cells and we now report that the cloned LcUSP is functional in vivo and can act as a partner for a chimeric L. cuprina ecdysone receptor to form an ecdysteroid-dependent transcription factor in mammalian cells.
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PMID:LcUSP, an ultraspiracle gene from the sheep blowfly, Lucilia cuprina: cDNA cloning, developmental expression of RNA and confirmation of function. 1137 12

Protein-protein interactions are critical for the function of biological systems. Here, we describe a means to dissect a protein-protein interaction. Our method is based on the in vivo interaction between a target protein and the peptide epitopes derived from its partner. This interaction is detected by using hybrid proteins in which the target protein and peptide epitopes are fused to the DNA-binding domain of the lambda repressor protein. An interaction prevents the transcription of a reporter gene. The efficacy of this approach is demonstrated with the ribonuclease inhibitor protein and ribonuclease A, which form a complex with an equilibrium dissociation constant in the femtomolar range. Our method can enable the identification of residues important in a designated protein-protein interaction and the development of antagonists for that interaction.
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PMID:Genetic screen to dissect protein-protein interactions: ribonuclease inhibitor-ribonuclease A as a model system. 1243 38