Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.30.1 (S1 nuclease)
 3,660 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In a previous publication (Wen, L.-P., Ruettinger, R. T., and Fulco, A.J. (1989) J. Biol. Chem. 264, 10996-11003), we reported that about 1 kilobase of 5' flanking sequence was required for barbiturate-inducible expression of the cytochrome P450BM-3 gene in Bacillus megaterium. We have now found, by analysis of various deletion and frameshift derivatives of this region, that an open reading frame immediately upstream of the B. megaterium cytochrome P450BM-3 structural gene encodes a protein, designated Bm3R1, which negatively controls the expression of the P450BM-3 gene at the transcriptional level. A helix-turn-helix DNA binding motif was found near the N-terminal portion of Bm3R1 protein. The 5' terminus of the bm3R1 transcript generated in vivo was determined by nuclease S1 mapping and primer extension analysis to be 44 base pairs upstream of the translation initiation sequence GTG of bm3R1. A putative promoter sequence with a high degree of similarity to the -35 and -10 consensus sequence recognized by the Bacillus subtilis sigma-43 factor was located at an appropriate distance from the transcription start site. A B. megaterium mutant which highly constitutively produced P450BM-3 protein was isolated and complementation of this cytochrome P450BM-3-constitutive mutant by a DNA fragment containing the wild-type bm3R1 gene indicated that the mutation in this locus was trans-dominant. Sequence analysis of the bm3R1 gene and its upstream region from this mutant, after amplification by the polymerase chain reaction, identified a single base change that resulted in a glycine to glutamate substitution in the beta-turn region of the DNA binding motif. By placing the bm3R1 gene under the control of a tac promoter and changing the translation initiation sequence from GTG to ATG, we succeeded in overproducing the Bm3R1 protein in Escherichia coli. A 20-bp perfect palindromic putative operator site, located between the presumed promoter sequences and the bm3R1 structural gene, was defined both by in vivo titration of Bm3R1 repressor and by gel mobility shift assays using the cell-free extracts containing the overproduced wild-type or mutant Bm3R1 protein. The barbiturate effect in mediating the induction of cytochrome P450BM-3 appears to be indirect but probably involves, in part, the release of inhibition by Bm3R1 repressor protein by interfering with its binding to the palindromic putative operator sequence and perhaps to other sites on the regulatory region of the gene encoding cytochrome P450BM-3.
 ...
PMID:Barbiturate-mediated regulation of expression of the cytochrome P450BM-3 gene of Bacillus megaterium by Bm3R1 protein. 154 26

Giardia lamblia is believed to be the earliest branching derivative from the eucaryotic lineage. Genomic and cDNA clones encoding the giardia NADP-dependent glutamate dehydrogenase have been isolated and characterized. Southern hydridization using genomic DNA indicates that the gene encoding this activity is unique and single copy. Primer extension, S1 nuclease protection, and genomic and cDNA sequence analysis demonstrate that gene transcripts are initiated within a conserved AT-rich sequence element immediately preceding the ATG translation initiation codon and the short 5' untranslated region is not extended by transsplicing. The open reading frame is 1350 nucleotides in length and encodes a protein of 449 amino acids. The reading frame is not interrupted by introns and the primary transcript is probably not subjected to RNA editing. In the strictly anaerobic metabolism of giardia, NADP-dependent glutamate dehydrogenase activity participates along with alanine aminotransferase, in the cyclic dissipation of reducing equivalents (NADPH) through the conversion of pyruvate to alanine. The deduced amino acid sequence of the giardia protein exhibits substantial homology to numerous fungal and eubacterial NADP-dependent glutamate dehydrogenases. Comparisons of alignment gap positions and amino acid identities indicate that the giardia sequence is at least as similar or more similar to the eubacterial sequence than it is to the fungal sequence. This supports the hypothesis that giardia diverged very early from the eucaryotic lineage.
 ...
PMID:Isolation and characterization of a NADP-dependent glutamate dehydrogenase gene from the primitive eucaryote Giardia lamblia. 155 91

The yeast GDH1 gene encodes NADP-dependent glutamate dehydrogenase. This gene was isolated by complementation of an Escherichia coli glutamate auxotroph. NADP-dependent glutamate dehydrogenase was overproduced 6-10-fold in Saccharomyces cerevisiae bearing GDH1 on a multicopy plasmid. The nucleotide sequence of the 1362-base pair coding region and 5' and 3' flanking sequences were determined. Transcription start sites were located by S1 nuclease mapping. Regulation of GDH1 was not maintained when the gene was present on a multicopy plasmid. Protein secondary structure predictions identified a region with potential to form the dinucleotide-binding domain. The amino acid sequences of the yeast and Neurospora crassa enzymes are 63% conserved. Unlike the N. crassa gene, yeast GDH1 has no introns.
 ...
PMID:Nucleotide sequence of yeast GDH1 encoding nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase. 298 90

Type IV pili are required for social gliding motility in Myxococcus xanthus. In this work, the expression of pilin (the pilA gene product) during vegetative growth and fruiting-body development was examined. A polyclonal antibody against the pilA gene product (prepilin) was prepared, along with a pilA-lacZ fusion, and was used to assay expression of pilA in M. xanthus in different mutant backgrounds. pilA expression required the response regulator pilR but was negatively regulated by the putative sensor kinase pilS. pilA expression did not require pilB, pilC, or pilT. pilA was also autoregulated; a mutation which altered an invariant glutamate five residues from the presumed prepilin processing site eliminated this autoregulation, as did a deletion of the pilA gene. Primer extension and S1 nuclease analysis identified a sigma54 promoter upstream of pilA, consistent with the homology of pilR to the NtrC family of response regulators. Expression of pilA was found to be developmentally regulated; however, the timing of this expression pattern was not entirely dependent on pilS or pilR. Finally, pilA expression was induced by high nutrient concentrations, an effect that was also not dependent on pilS or pilR.
 ...
PMID:Regulation of expression of the pilA gene in Myxococcus xanthus. 940 Oct 34

The complete nucleotide sequence for the aot operon of Pseudomonas aeruginosa PAO1 was determined. This operon contains six open reading frames. The derived sequences for four of these, aotJ, aotQ, aotM, and aotP, show high similarity to those of components of the periplasmic binding protein-dependent ABC (ATP binding cassette) transporters of enteric bacteria. Transport studies with deletion derivatives established that these four genes function in arginine-inducible uptake of arginine and ornithine but not lysine. The aotO gene, which encodes a polypeptide with no significant similarity to any known proteins, is not essential for arginine and ornithine uptake. The sixth and terminal gene in the operon encodes ArgR, which has been recently shown to function in regulation of arginine metabolism. Studies with an aotJ::lacZ translational fusion showed that expression of the aot operon is strongly induced by arginine and that this effect is mediated by ArgR. S1 nuclease and primer extension experiments showed the presence of two promoters, P1 and P2. The downstream promoter, P2, is induced by arginine and appears to be subject to carbon catabolite repression. The upstream promoter, P1, is induced by glutamate. Footprinting experiments established the presence of a 44-bp ArgR binding site that overlaps the -35 region for P2, as was shown to be the case for the arginine-inducible aru promoter, and the -10 region for P1, as was shown to be the case for arginine-repressible operons in P. aeruginosa. Sequence alignment confirms the architecture and the consensus sequence of the ArgR binding sites, as was previously reported.
 ...
PMID:Molecular characterization and regulation of an operon encoding a system for transport of arginine and ornithine and the ArgR regulatory protein in Pseudomonas aeruginosa. 979 Nov 3