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  • 1
    ISSN: 1432-2048
    Keywords: Key words: Abscisic acid ; Gene expression (organ specific) ; Histidine-rich duplicated domain ; Oryza (salt stress) ; Salt stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. A novel cDNA clone osr40c1, encoding a abscisic acid (ABA)-responsive 40-kDa protein previously associated with salt tolerance (Moons et al. `1995' Plant Physiol 107: 177–186), was isolated from roots of rice seedlings (Oryza sativa L.). Exogenously applied ABA and salt shock induced a marked increase of the osr40c1 transcript level in roots of seedlings whereas constant osr40c1 mRNA levels were found in the shoot. The root-specific salinity-induced osr40c1 mRNA accumulation was rapid and gradually declined upon prolonged salt shock. Plant growth regulators, signalling the wounding and the pathogen response, did not enhance osr40c1 expression, indicating a salt- and osmotic-stress-specific response. The encoded OSR40c1 protein was found to be hydrophilic, rich in histidine residues (6%) constituting putative metal-binding domains, and to consist of a duplicated domain of 151 amino acids (75% identical), that can form amphiphilic α-helical structures. The gene osr40c1 belongs to a multigene family. Two osr40 genes were isolated, osr40g2 and osr40g3, tandemly arranged in an 8-kb region of the rice genome. Antisera raised against a conserved OSR40 peptide recognized different OSR40 proteins that accumulated in roots upon exposure to salt stress. The OSR40 protein family included 29-kDa proteins and two 40-kDa proteins, the latter most probably corresponding to OSR40c1 and OSR40g2 with duplicated domain structures. The osr40g3 transcript encoded a single copy of the OSR40 domain and exhibited a shoot-specific expression. Results indicate that OSR40c1 plays a role in the adaptative response of roots to an hyper-osmotic environment and belongs to a novel plant protein family that most probably has structural functions.
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  • 2
    ISSN: 1432-072X
    Keywords: Key words GroEL ; GroES ; Protein purification ; ATPase activity ; Chaperonin activity ; Heat-shock ; induction ; Heat-shock promoter mapping ; Rhodobacter capsulatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The heat-shock protein Cpn60 (chaperonin, GroEL homologue) from the phototrophic bacterium Rhodobacter capsulatus B10 was purified to homogeneity and biochemically characterized. Native Cpn60 from R. capsulatus was shown to be a tetradecamer of 840 kDa similar to that of homologous chaperones characterized so far. Cpn60 possesses ATPase activity and promotes refolding of chaotropically denatured citrate synthase. The groESL operon of R. capsulatus was cloned using a degenerate oligonucleotide and sequenced. Two open reading frames (285 and 1,635 bp) were found; they encode Cpn10 and Cpn60, with corresponding deduced molecular masses of 10.6 and 57.6 kDa. The deduced amino acid sequences coincided perfectly with those of the amino terminus and of three tryptic peptides of purified Cpn60 from R. capsulatus. Strong evidence that R. capsulatus encodes only one copy of the groESL operon was obtained. Primer-extension analysis revealed that the groESL operon is transcribed by a –35/–10-type promoter, and that transcription was initiated from the same positions before and after heat-shock under both chemotrophic and phototrophic conditions. The major initiation site is immediately followed by the inverted repeat structure CIRCE, which has been found upstream of many bacterial heat-shock operons. A second minor transcript starts just after the CIRCE element. Although heat-shock induction of a groEL-lacZ fusion failed because of thermal inactivation of the fusion protein, Western blot analysis revealed a two- to threefold induction of cellular Cpn60 levels 45–75 min after shifting from 28° C to 39° C. Deletion mapping of the groESL promoter identified upstream of the promoter a 19-bp element that enhances groESL transcription eightfold and contains the AT-rich sequence dAAATTTTT, which is found at similar positions in heat-shock operons of other gram-negative bacteria.
    Type of Medium: Electronic Resource
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