Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The pattern of lipoxygenase (LOX) gene expression was investigated in pea nodule tissues using the technique of in situ hybridization. Five lipoxygenase cDNAs were cloned from nodule mRNA by the RT-PCR and 3' RACE procedures. These clones (loxN1 to loxN5) show a high degree of sequence homology, except in the 3'-untranslated region. Gene-specific riboprobes were therefore generated from subclones carrying the 3'-untranslated regions in order to investigate tissue-specific gene expression. Northern blotting analysis revealed that loxN1 corresponded to a transcript that was expressed exclusively in roots and nodules but not in the aerial parts of the plant. However, none of the LOX genes appeared to be up-regulated in nodule tissue relative to uninfected roots. Starting with the incomplete cDNA clone for loxN1, the full coding sequence termed lox1:P.s:1 was obtained by further rounds of RT-PCR and 5' RACE procedures. In situ hybridization with nodule tissues revealed several different patterns of expression for the various LOX probes. However, none of the corresponding transcripts was expressed exclusively in the invasion zone, as might have been expected if one LOX gene product had been uniquely associated with the invasion process. In conclusion, this study provides no evidence for a direct role for any LOX gene product in plant-microbe interaction or host defence, but the fact that all the transcripts were expressed at the nodule apex suggests that LOX could be involved in the development of this organ.
Plant Mol Biol 1999 Mar
PMID:Isolation of lipoxygenase cDNA clones from pea nodule mRNA. 1035 91

The cell wall acts as the first line of defense during pathogen invasion. Polygalacturonases (PGs) are a class of cell-wall-modifying enzymes with precise temporal and organ-specific expression. A 350-bp fragment with high homology to PGs was identified by differential display (DD) analysis of soybean cyst nematode (SCN) race 3 resistant PI 437654 and susceptible cultivar Essex. The fragment was strongly expressed in Essex, 2 days after inoculation (DAI). Complete coding sequences of two PG cDNAs, PG1 and PG2, were isolated by 3' and 5' rapid amplification of cDNA ends polymerase chain reaction (RACE PCR). PI 437654 and Essex had identical PG1 and PG2 sequences. A transversion from A to C created a PstI restriction site in the PG2 cDNA that was used to distinguish the two PG cDNAs by cleaved amplified polymorphic sequence (CAPS) analysis. A cDNA encoding a polygalacturonase-inhibitor protein (PGIP) that is 89% identical to the Phaseolus vulgaris PGIP was isolated from soybean roots by reverse transcription (RT)-PCR. Steady-state levels of PG and PGIP were investigated by RNA gel blot analysis in roots 1 to 5 DAI and in hypocotyls and leaves. Differences in the constitutive levels of PG mRNAs were observed in roots of different soybean genotypes. Steady-state levels of PG mRNAs were enhanced during compatible interactions with SCN and reduced in incompatible interactions and in mechanically wounded roots. Enhanced PGIP transcription was observed in response to mechanical wounding in both PI 437654 and Essex, but only in compatible interactions with SCN, suggesting uncoupling of PGIP functions in developmental and stress cues. Constitutive expression in incompatible interactions shows PGIP is not a factor in SCN resistance. Thus, the up-regulation of endogenous PG transcription in soybean roots early after SCN infection could facilitate successful parasitism by SCN.
Mol Plant Microbe Interact 1999 Jun
PMID:Polygalacturonase and polygalacturonase inhibitor protein: gene isolation and transcription in Glycine max-Heterodera glycines interactions. 1035

Utrophin is a large protein which accumulates at the neuromuscular synapse and myotendinous junctions in adult skeletal muscle, and is widely expressed in several non-skeletal muscle tissues. Evidence from a variety of sources suggests that a successful strategy for treatment of Duchenne muscular dystrophy patients will be to increase expression of utrophin in muscle. There is still much to be learnt about utrophin gene regulation, in particular regarding alternative isoforms, their promoters and role in muscle and non-muscle tissues. Using 5"-RACE we have identified two novel transcripts of utrophin, Up71 and Up140, with unique first exons and promoters located in intron 62 and intron 44, respectively. These transcripts appear to be structural homologues of the short dystrophin transcripts, Dp140 and Dp71, emphasizing the high degree of structural conservation between the utrophin and dystrophin genes. RT-PCR shows that Up71 and Up140 are widely expressed in both human and mouse tissues, including skeletal muscle. We present evidence for transcript-specific differential mRNA splicing of exon 71, in both Up71 and Up140, similar to that described for dystrophin. No evidence for splicing of exon 78 of utrophin was found. This is in contrast to dystrophin and may reflect a subtle functional difference in patterns of phosphorylation between the two proteins.
Hum Mol Genet 1999 Jul
PMID:Up71 and up140, two novel transcripts of utrophin that are homologues of short forms of dystrophin. 1036 73

Acyl-CoA delta 9-desaturases play essential roles in fatty acid metabolism and the regulation of cell membrane fluidity. In this research, a cDNA sequence was obtained from Trichoplusia ni adult fat body mRNA by using RT-PCR with degenerate primers based on other characterized delta 9-desaturase sequences. The remainder of the sequence was amplified using 3'- and 5'-RACE. A 1439 bp cDNA reconstructed from three overlapping PCR products contains an ORF encoding a 353-amino acids (aa) protein that shows clear homology (greater than 50% aa identity and greater than 65% aa similarity to characterized insect and vertebrate desaturases). The ORF of this cDNA was subcloned into an expression vector, which relieved the unsaturated fatty acid (UFA) auxotrophy of a desaturase-deficient yeast strain following genetic transformation. The newly characterized desaturase from T. ni produced fatty acids delta 9-16 and delta 9-18 in a 1:6 ratio, compared to a 5:1 ratio, respectively, with the yeast delta 9 desaturase. A Northern blot hybridization and a RT-PCR experiment showed that temporal and tissue-specific patterns of expression of the corresponding mRNA are distinct from those of the delta 11-desaturase mRNA present in the pheromone glands of adult females. Based on its homology to other desaturases, the widespread distribution of its corresponding mRNA in various tissues, and its functional assay, we conclude that this cDNA encodes the apoprotein corresponding to the desaturase component of the metabolic delta 9-desaturase complex of T. ni.
Insect Biochem Mol Biol 1999 May
PMID:Cloning and functional expression of a cDNA encoding a metabolic acyl-CoA delta 9-desaturase of the cabbage looper moth, Trichoplusia ni. 1038 Jun 55

The regulation of mu-opioid receptor gene expression was investigated using several molecular techniques. Genomic clones containing portions of the human mu-opioid receptor gene were sequenced. 5'-RACE analysis of human brain cDNA confirmed the presence of mRNAs up to -313 from the start codon. As was found for the mouse and rat genes, transcription apparently initiates in the absence of a discernable TATA box. To characterize promoter function, portions of the 5'-flanking region were linked to a reporter gene in transient transfection experiments. Two approximately 50 bp adjacent segments had potent, orientation specific promoter activity. More down-stream segments also had promoter activity. None of the 5'-flanking region constructs showed tissue specificity. The potential role of DNA methylation in preventing ectopic expression was investigated by surveying the methylation state of a CpG rich region straddling the start codon. A neural derived cell line (SH-SY5Y) that expresses the mu-opioid receptor lacked virtually any CpG methylation. In contrast, two neural derived cell lines that do not express the mu-opioid receptor were nearly totally methylated while non-neural cell lines had intermediate levels of CpG methylation. Additional transient transfection experiments revealed that CpG methylation of the 5'-flanking region suppressed reporter gene expression. These results indicate that CpG methylation plays an important role in regulating mu-opioid receptor expression in neural cells; however, no association was found with regulation of expression in non-neural cells.
Brain Res Mol Brain Res 1999 Jun 18
PMID:Localization of promoter elements in the human mu-opioid receptor gene and regulation by DNA methylation. 1038 43

Differential display PCR was used to identify genes regulated by mood stabilizer lithium in rat cerebral cortex. A differentially displayed lithium regulated gene fragment was isolated in rat cerebral cortex after chronic treatment with lithium (1.69 g/kg, p.o. ) for three weeks. A 1216-nucleotide cDNA for a novel lithium regulated gene (NLRG) was isolated from a rat brain cDNA library with RACE (rapid amplification of 5' cDNA end) PCR using a prime from the differentially displayed NLRG gene fragment. The deduced protein sequence was 321 amino acids long, and shows a significant homology with yeast nitrogen permease regulator 2 (NPR2). NLRG expression induced by lithium was confirmed by Northern and slot blot analysis in rat cerebral cortex and neuroblastomaxglioma NG108-15 cells, respectively. In situ hybridization revealed that chronic treatment with lithium increased NLRG gene expression in frontal cortex and hippocampus, but not in striatum, hypothalamus and thalamus regions of rat brain. These results suggest a novel target for lithium which may be relevant to its mechanism of action.
Brain Res Mol Brain Res 1999 Jun 18
PMID:Identification of a novel lithium regulated gene in rat brain. 1038 44

Growth hormone (GH) gene expression has been reported in the mammary glands of various mammalian species. The mechanism by which the GH gene becomes activated in extrapituitary tissues is currently unclear. We have characterized the canine mammary and pituitary GH gene transcripts by Northern blot, 5'- and 3'-RACE (rapid amplification of cDNA ends), and DNA sequence analysis. Northern blot analysis detected GH gene transcripts in mammary glands of dogs which were exposed to high levels of progestins. The mammary and pituitary GH cDNAs were shown to be identical in both the coding region and untranslated regions. Pituitary GH gene expression is highly dependent upon the transcription factor Pit-1. Analysis of Pit-1 gene expression using RT-PCR followed by Southern hybridization revealed a strong pituitary signal but faint, weak or no hybridization signals in mammary gland samples. Among the negative samples were progestin-treated dogs with high mammary GH gene expression. These findings indicate that mammary and pituitary GH gene transcripts originate from the same transcription start site but are regulated differentially.
Mol Cell Endocrinol 1999 Apr 25
PMID:Canine mammary growth hormone gene transcription initiates at the pituitary-specific start site in the absence of Pit-1. 1041 6

Plasmodium merozoites are covered with a palisade layer of proteins that are arranged as organized bundles or appear as protruding spikes by electron microscopy. Here we present a third Plasmodium vivax merozoite surface protein, PvMSP-3, which is associated with but not anchored in the merozoite membrane. Serum from a P. vivax immune squirrel monkey was used to screen a lambdagt11 P. vivax genomic DNA (gDNA) library. Plaque-selected antibodies from clone no. 6.1, and rabbit antisera against its encoded protein, produced a pattern in immunofluorescence assays (IFAs) that is consistent with a localization at the surface of mature schizonts and free merozoites. Specific antisera also agglutinated merozoites and recognized a protein of 150 000 Da by SDS-PAGE. The complete msp-3 gene and flanking sequences were cloned from a P. vivax lambda Dash II gDNA library and also partly characterized by RACE (rapid amplification of cDNA ends). The immediate upstream sequence contains non-coding repeats and a putative protein encoding open reading frame (ORF), which are also present on the msp-3 5'RACE gene product. Pvmsp-3 encodes a protein with a calculated mass of 89 573 Da, which has a potential signal peptide and a major central alanine-rich domain (31%) that exhibits largely alpha-helical secondary structure and is flanked by charged regions. The protein does not have a putative transmembrane domain or a consensus sequence for a glycosylphosphatidylinositol (GPI) anchor modification. However, the alanine-rich domain has heptad repeats that are predicted to form coiled-coil tertiary structures, which mediate protein-protein interactions. PvMSP-3 is structurally related to P. falciparum MSP-3 and the 140000 Da MSP of P. knowlesi. Characterization of PvMSP-3, thus, also begins to define a new interspecies family of evolutionarily related Plasmodium merozoite proteins.
Mol Biochem Parasitol 1999 Jun 25
PMID:Plasmodium vivax merozoite surface protein-3 contains coiled-coil motifs in an alanine-rich central domain. 1041 49

The straw mushroom Volvariella volvacea is cultivated on substrates rich in cellulose and has been shown to produce a family of cellulolytic enzymes. A PCR-based strategy was adopted to clone genes involved in cellulose utilisation, using degenerate primers designed to amplify conserved catalytic domain sequences of cellobiohydrolases (CBHs). PCR with these primers produced two DNA fragments with sequence similarity to the cbhI and cbhII gene families detected in Trichoderma, Phanerochaete and Agaricus species. Full-length clones of these genes were obtained from an EMBL3 genomic library, and RACE-PCR was used to verify the presence of introns. The cbhI homologue has a coding region of 1722 bp, containing two introns, generating a 536 amino acid polypeptide product. The cbhII gene has a coding region of 1693 bp, containing five introns, and gives rise to a 470-amino acid polypeptide product. Northern and PCR analyses were used to study the expression of the genes. These revealed that transcripts of both genes were induced on medium containing cellulose with cbhI being expressed more strongly than cbhII - but were repressed on medium containing glucose.
Mol Gen Genet 1999 Jul
PMID:Cloning of the cbhI and cbhII genes involved in cellulose utilisation by the straw mushroom Volvariella volvacea. 1048 90

We have cloned, characterized and sequenced the murine TNF Receptor Associated Factor 1 (TRAF1) gene. Restriction mapping and Southern blotting analysis revealed that the TRAF1 gene comprises 10 exons and 9 intervening introns and spreads over 18 kb of genomic DNA. 5'-RACE analysis of the TRAF1 transcript using mRNA from activated spleen B cells revealed several transcription start sites between positions -42 to +4 relative to the 5'end of the murine TRAF1 cDNA sequence. We also isolated and sequenced the 5'-upstream promoter region, which lacks TATA-like and CAAT-like sites but contains GC-rich sequences. Taken together, these results suggest that the TRAF1 gene promoter is a member of the class of Sp-1-dependent promoters. Near the transcription initiation start site we identified three identical decanucleotide repeats (CCAGCCCAGC) which may play a role in the transcriptional regulation of TRAF1 expression. In addition we show that TRAF1 mRNA is not expressed in non-stimulated lymphocytes but can be induced upon activation with different stimuli, including anti-CD3, anti-IgM, anti-CD40 antibodies, LPS, or a combination of phorbol-12-myristate-13-acetate and ionomycin.
Mol Immunol 1999 Jun
PMID:Structure of the murine TRAF1 gene. 1049 14


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