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Query: UNIPROT:P06889 (Mol)
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Cold storage of potato tubers at 4 degrees C is associated with the accumulation of several cold-induced transcripts. By using a previously characterized cDNA (CI7) as probe, we isolated and sequenced the corresponding ci7 gene. The putative promoter of ci7 contains sequence elements that have been shown to mediate expression of stress-responsive genes of Arabidopsis thaliana. CI7 transcripts were differentially induced in response to cold, drought, high salt or exogenous ABA treatment in potato tubers and leaves. Whereas accumulation of CI7 transcript during cold storage occurred within days, induction of CI7 transcript in response to drought, ABA and salt occurred rapidly within few hours. In tubers, accumulation of CI7 protein in response to abiotic stresses and ABA was small when compared to transcript levels. In leaves, the CI7 protein was undetectable after all treatments tested. 3 kb of the 5'-flanking ci7 promoter region were fused to the GUS reporter gene and introduced into S. tuberosum plants. The analysis of tubers of independent transgenic lines did not reveal significant induction of enzymatic GUS activity in response to low temperature. When RNA gel blotting was used to analyze the level of induction of the GUS gene driven by the ci7 promoter, the heterologous GUS fusion was, however, strongly responsive to low temperature. Nuclear run-on transcription studies of the ci7 gene, in comparison with RNA gel blot analyses of the transgenic plants, indicated that most of the temperature-regulated expression of the ci7 gene in tubers may be accounted for by post-transcriptional control mechanisms.
Plant Mol Biol 1997 Mar
PMID:Structural organization, expression and promoter activity of a cold-stress-inducible gene of potato (Solanum tuberosum L.) 910 13

Excitotoxic amino acids, such as glutamate, may play an important role in retinal ischemia/reperfusion damage. In central neurons, excitotoxicity may be mediated by nitric oxide synthase (NOS) causing DNA damage via nitric oxide (NO). The nicked DNA activates poly-adenosine diphosphate (ADP)-ribose polymerase (PARP) and may deplete intracellular ATP resulting in cell death. PARP may also be involved in apoptosis. We used 3-aminobenzamide (3-ABA), a PARP inhibitor, to examine the possible involvement of PARP in a rat model of retinal ischemia. Retinal ischemia was induced by elevating the intraocular pressure (IOP) through the insertion of a needle into the anterior chamber of a rat eye. IOP was raised to 110 mm Hg for 60 minutes. Animals were given intracameral infusion of 0, 1, 3, 10, 30, 100 mM 3-ABA in 0.1 M PBS, pH 7.4 during ischemia. Morphologic and morphometric evaluation at 7 days after reperfusion showed that 3-ABA at 3 mM and above significantly ameliorated the ischemic/reperfusion damage to the retina. In addition, at 10 mM 3-ABA inhibited the characteristic ladder pattern in DNA gel analysis seen in apoptosis of retinal neurons after ischemia/reperfusion. Hence, PARP may be involved in retinal cell loss after ischemia/reperfusion insult probably through the apoptotic pathway.
Res Commun Mol Pathol Pharmacol 1997 Mar
PMID:The effect of 3-aminobenzamide, an inhibitor of poly-ADP-ribose polymerase, on ischemia/reperfusion damage in rat retina. 914 32

Maize root membranes were extracted and the solubilized proteins were affinity-purified using an ABA-BSA-Sepharose 4B matrix. The retained proteins were eluted with 4M urea or 10(-4)M ABA. ABA could elute the binding proteins but other phytohormones, such as IAA or GA3, could not. ABA binding activity was detected in ABA- and urea-elute fractions using competitive ELISA block tests and [3H]ABA binding assays. Scatchard analysis showed an apparent K(d) of 4.8 nM for the ABA binding activity of the protein. When ABA or urea eluate was loaded on a concanavalin-A-Sepharose column, the fraction eluted with 0.2 M methyl alpha-mannopyranoside still showed ABA binding activity, suggesting that ABA binding proteins are glycoproteins. Polyclonal antibodies against ABA binding proteins were raised using as immunogen ABA or urea eluate from the ABA-BSA-Sepharose column. The resulting antibodies not only recognized 56 kDa binding proteins but also blocked the binding of ABA to an ABA-specific antibody, indicating properties similar to anti-idiotypic antibodies. The purified antibodies will be suitable to purify and characterize putative ABA receptors.
J Mol Recognit
PMID:Purification and identification of ABA-binding proteins and antibody preparation. 917 63

To isolate genes which are expressed preferentially during embryogenesis, a Douglas-fir embryogenesis cDNA library was constructed and differentially screened with cDNA probes made with mRNA from developing and mature embryos, respectively. The cDNA clone PM 2.1 was isolated based on its abundance in developing seeds and absence in mature seeds, and its predicted amino acid sequence was shown to have structural features characteristic of plant MT-like proteins. Alignment of the PM 2.1 predicted amino acid sequence with other plant MT-like protein sequences revealed a general paucity of Cys and Cys-Xaa-Cys sequences and the presence of novel serine residues within the conserved Cys-Xaa-Cys motifs in the C-terminal domain. The consensus sequence following the Cys-poor spacer in type 2 MT-like proteins, CXCXXXCXCXXCXCX, was modified in PM 2.1 to CXSXXXSXYXX-XCX. Phylogenetic analysis supported PM 2.1 was distinct from other MT and grouped with MT-like proteins from Arabidopsis (OEST), rice (AEST) and kiwifruit (AD1), which do not belong to type 1 or 2. The PM 2.1 gene was expressed in somatic and zygotic embryos, in haploid maternal tissue, as well as in hormone- and metal-treated seeds and seedlings. The PM 2.1 transcripts were detected in the needles of 14-week-old seedlings, but not the root tissue or mature pollen. The expression of the PM 2.1 gene in embryos was dependent upon ABA and osmoticum and in seedlings was differentially modulated by metals, suggesting a role of the PM 2.1 gene product in the control of microelement availability during Douglas-fir seed development and germination. The novel structural features, and the developmental, hormonal and metal modulation of PM 2.1 expression, are evidence for a new type of MT-related protein in plants.
Plant Mol Biol 1997 May
PMID:The isolation of a novel metallothionein-related cDNA expressed in somatic and zygotic embryos of Douglas-fir: regulation by ABA, osmoticum, and metal ions. 920 40

The low-temperature (2 degrees C)-specific wheat cDNA, pTACR7, represents a gene designated tacr7 from hard red winter wheat (HRWW; Triticum aestivum L. cv. Winoka). The term low-temperature-specific (LTS) is used because tacr7 is not induced by ABA or stresses such as salt, dehydration, and heat. pTACR7 was isolated by RT-PCR with mRNA from wheat crown tissue, the oligonucleotide primers derived from the barley cognate pHVCR8 (GenBank accession number L28091). Based on the deduced amino acid sequence, TACR7 is highly hydrophobic, with a single transmembrane domain and an amino acid bias for leucine (19%). Thus, the encoded protein TACR7 is unique among low-temperature-regulated wheat proteins described in the literature. Analysis of steady-state levels of tacr7 transcripts (630 nt) showed accumulation in wheat seedlings, crown tissue, and callus cultures after transfer from control (25 degrees C) to low temperature (2 degrees C). No detectable transcripts were observed by northern blot hybridization with pTACR7 probe from seedling or callus treated with ABA, salt, dehydration, or heat stress. tacr7 transcripts accumulated during 2 degrees C exposure to a greater amount in a freeze-resistant HRWW (FR; SDmut 16029) than in a freeze-susceptible HRWW (FS; SDmut 16169) crown tissue, with the largest difference between genotypes being 30% +/- 3% at 3 weeks.
Plant Mol Biol 1997 Jul
PMID:cDNA structure and expression patterns of a low-temperature-specific wheat gene tacr7. 924 45

Six cDNA clones from Phaseolus vulgaris, whose expression is induced by water deficit and ABA treatment (rsP cDNAs) were identified and characterized. The sequence analyses of the isolated clones suggest that they encode two types of late-embryogenesis abundant (LEA) proteins, a class-1 cytoplasmic low-molecular-weight heat shock protein (lmw-HSP), a lipid transfer protein (LTP), and two different proline-rich proteins (PRP). One of the putative LEA proteins identified corresponds to a novel 9.3 kDa LEA-like protein. During the plant response to a mild water deficit (psi w = -0.35 MPa) all genes identified present a maximal expression at around 16 or 24 h of treatment, followed by a decline in expression levels. Rehydration experiments revealed that those genes encoding PRPs and LTP transiently re-induce or maintain their expression when water is added to the soil after a dehydration period. This is not the case for the lea genes whose transcripts rapidly decrease, reaching basal levels a few hours after rehydration (4 h). Under water deficit and ABA treatments, the highest levels of expression for most of the genes occur in the root, excluding the ltp gene whose maximum expression levels are found in the aerial regions of the plant. This indicates that for these genes, both water deficit and ABA-dependent expression are under organ-specific control. The data presented here support the importance of these proteins during the plant response to water deficit.
Plant Mol Biol 1997 Nov
PMID:Characterization of Phaseolus vulgaris cDNA clones responsive to water deficit: identification of a novel late embryogenesis abundant-like protein. 934 63

The transcription factors VP1 (Viviparous-1), EmBP-1 (Em-binding protein 1) and OSBZ8, originally cloned and analysed in various monocot species, have been implicated in the regulation of the Lea (late embryogenesis-abundant) group 1 genes. We have investigated the modulation of the levels of these mRNAs in barley during embryogenesis, and in developing embryos subjected to various kinds of osmotic stress. The accumulation of mRNA for VP1 and EmBP-1 transcription factors, using cDNAs cloned from barley, starts at 10 and 15 days after anthesis, respectively, whereas Lea B19 mRNA levels are very low or undetectable until 25 days after anthesis during normal development. The EmBP-1 mRNA is predominantly induced in mannitol-stressed immature embryos. Vp1 mRNA was not significantly modulated by ABA, salt or mannitol. Inhibition of ABA biosynthesis by norflurazon showed that the induction of both Vp1 and EmBP-1 mRNAs was ABA-independent. In embryo-derived suspension-cultured cells, neither of the two transcripts would be induced by ABA or osmotic stress, although both OSBZ8 and one member of the Lea B19 family was up-regulated by ABA. Electrophoretic mobility shift assays using a Lea B19.1 probe with an ABRE (abscisic acid-responsive element) similar to that which binds EmBP-1 and OSBZ8 in the wheat and rice Em promoters show that the binding activity is increased by ABA and osmotic stress. Taken together, these data show that both VP1 and EmBP-1 are involved in embryo-specific signal transduction pathways, that they are differentially regulated at the mRNA level, and that EmBP-1 can be induced by osmotic stress independently of any increase in endogenous ABA. The difference in mRNA regulation patterns of OSBZ8 and EmBP-1 may suggest that they are involved in different signal transduction pathways in connection with osmotic stress/ABA regulation of Lea genes.
Plant Mol Biol 1997 Nov
PMID:Developmental, stress and ABA modulation of mRNA levels for bZip transcription factors and Vp1 in barley embryos and embryo-derived suspension cultures. 934 78

A cDNA clone (lp3) from loblolly pine induced by water deficit stress (WDS) has been isolated. It is preferentially induced in roots with a constitutive basal level of expression also observed in stems and needles. Northern blot analysis with well irrigated ABA-treated seedlings indicated that the overall accumulation of lp3 transcripts in the roots was lower than that of water deficit-stressed seedlings. However, within roots, lp3 was induced by ABA indicating that the expression of lp3 in roots under WDS conditions was partly mediated by ABA. The lp3 clone is similar to a group of genes called asr (ABA stress and ripening) genes identified in several species. A genomic clone (lp3-1) was identified and its putative protein has the hydrophylicity profile similar to that of lp3 except for two deletions in the 5' region. The genomic Southern and RT-PCR (reverse transcriptase-polymerase chain reaction) analyses indicate that the lp3 gene belongs to a small multigene family of at least four members with a distinct pattern of expression during WDS.
Plant Mol Biol 1997 Dec
PMID:Expression analysis of a gene family in loblolly pine (Pinus taeda L.) induced by water deficit stress. 942

We report the isolation by differential display of a novel tomato ethylene-responsive cDNA, designated ER5. RT-PCR analysis of ER5 expression revealed an early (15 min) and transient induction by ethylene in tomato fruit, leaves and roots. ER5 mRNA accumulated during 2 h of ethylene treatment and thereafter underwent a dramatic decline leading to undetectable expression after 5 h of treatment. The full-length cDNA clone of 748 bp was obtained and DNA sequence analysis showed strong homologies to members of the atypical hydrophobic group of the LEA protein family. The predicted amino acid sequence shows 67%, 64%, 64%, and 61% sequence identity with the tomato Lemmi9, soybean D95-4, cotton Lea14-A, and resurrection plant pcC27-45 gene products, respectively. As with the other members of this group, ER5 encodes a predominantly hydrophobic protein. Prolonged drought stress stimulates ER5 expression in leaves and roots, while ABA induction of this ethylene-responsive clone is confined to the leaves. The use of 1-MCP, an inhibitor of ethylene action, indicates that the drought induction of ER5 is ethylene-mediated in tomato roots. Finally, wounding stimulates ER5 mRNA accumulation in leaves and roots. Among the Lea gene family this novel clone is the first to display an ethylene-regulated expression.
Plant Mol Biol 1997 Dec
PMID:ER5, a tomato cDNA encoding an ethylene-responsive LEA-like protein: characterization and expression in response to drought, ABA and wounding. 942 4

A cDNA clone was selected from a cDNA library constructed using mRNA from ABA-treated Fagus sylvatica L dormant seeds as a template. The clone is highly expressed in the presence of ABA and tends to disappear in stratified seeds. A search of sequence databases showed that the clone encodes a small GTP-binding protein. By means of in situ hybridization, the mRNA has been located in the apical meristem of the embryonic axis and in the central vascular cylinder. Its possible involvement in growth regulation in the embryonic axis of F. sylvatica is discussed.
Plant Mol Biol 1998 Feb
PMID:Transcripts of a gene, encoding a small GTP-binding protein from Fagus sylvatica, are induced by ABA and accumulated in the embryonic axis of dormant seeds. 948 89


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