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Query: UNIPROT:P06889 (Mol)
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The egg jelly (EJ) coat which surrounds the unfertilized sea urchin egg undergoes extensive swelling upon contact with sea water, forming a three-dimensional network of interconnected fibers extending nearly 50 microns from the egg surface. Owing to its solubility, this coat has been difficult to visualize by light and electron microscopy. However, Lytechinus pictus EJ coats remain intact, if the fixation medium is maintained at pH 9. The addition of alcian blue during the final dehydration step of sample preparation stains the EJ for visualization of resin embedded eggs by both light and electron microscopy. Stereo pairs taken of thick sections prepared for intermediate voltage electron microscopy (IVEM) produce a three-dimensional image of the EJ network, consisting of interconnected fibers decorated along their length by globular jelly components. Using scanning electron microscopy (SEM), we have shown that before swelling, EJ exists in a tightly bound network of jelly fibers, 50-60 nm in diameter. In contrast, swollen EJ consists of a greatly extended network whose fibrous components measure 10 to 30 nm in diameter. High resolution stereo images of hydrated jelly produced by the quick-freeze/deep-etch/rotary-shadowing technique (QF/DE/RS) show nearly identical EJ networks, suggesting that dehydration does not markedly alter the structure of this extracellular matrix.
Mol Reprod Dev 1993 Jun
PMID:The sea urchin egg jelly coat is a three-dimensional fibrous network as seen by intermediate voltage electron microscopy and deep etching analysis. 831 23

A cDNA clone, pMA1949, detects two mRNA species in wheat seedling tissue that are late embryogenesis-abundant (LEA) and dehydration stress-inducible. Sequence analysis of the pMA1949 clone shows it to be a 991 bp partial cDNA encoding a polypeptide of 317 amino acids with homology to two group 3 LEA proteins, carrot (DC8) and a soybean protein encoded by pGmPM2 cDNA. Molecular analysis of the deduced protein reveals a 33 kDa acidic and extremely hydrophilic protein with potential amphiphilic alpha-helical regions. In addition, the protein contains eleven similar, contiguous repeats of 11 amino acids, which are separated by 118 amino acids from two additional and unique repeats of 36 residues each at the carboxyl end of the protein. Comparisons of sequences of reported group 3 LEA proteins revealed that there are two types, separable by sequence similarity of the 11 amino acid repeating motifs and by the presence or absence of a certain amino acid stretch at the carboxyl terminus. Based on results from these comparisons, we propose a second type of group 3 LEA proteins, called group 3 LEA (II).
Plant Mol Biol 1993 Mar
PMID:Unusual sequence of group 3 LEA (II) mRNA inducible by dehydration stress in wheat. 846 82

Studies on vasoactive intestinal peptide (VIP) in the anterior pituitary gland have shown that it is synthesized locally, physiologically regulated, and may act as a paracrine/autocrine factor. We have now investigated the regulation of anterior pituitary VIP gene expression in rats during osmotic stimulation. Both salt-loading and dehydration resulted in a progressive and marked reduction in VIP mRNA levels as determined by Northern analysis, to 10% of control levels at 14 days of salt-loading. The 1.7 and 1.0 kb VIP RNA transcripts were equally affected. Since anterior pituitary VIP is partially localized in lactotrophs we also measured prolactin (PRL) mRNA levels. In contrast to VIP, PRL mRNA levels were increased during both osmotic paradigms, the mRNA levels being significantly raised after 5 days of salt-loading to 130% of controls. Further experiments, conducted to examine the mechanism by which VIP gene expression is down-regulated during osmotic stimulation, demonstrated that dopamine and angiotensin II do not appear to be involved. The results show dissociated regulation of VIP and PRL during osmotic stimulation and provide suggestive evidence of a role for anterior pituitary VIP in the animal's osmoregulatory responses. VIP may therefore be a paracrine factor with diverse functional roles.
Mol Cell Endocrinol 1993 Mar
PMID:Osmotic stimuli attenuate vasoactive intestinal peptide gene expression in the rat anterior pituitary gland. 847 72

Nine cDNA clones, corresponding to genes that are responsive to dehydration (named RD), have been isolated from Arabidopsis thaliana. The sequence of a putative protein encoded by one of the RD cDNA clones, RD22, exhibits considerable homology to an unidentified seed protein (USP) of Vicia faba. Northern analysis showed that RD22 mRNA is induced by salt stress as well as by water deficit but not by cold or heat stress. RD22 mRNA appeared after the application of abscisic acid (ABA), an indication that transcription of RD22 mRNA is induced by endogenous ABA, the production of which is triggered by drought and salt stress. The induction of RD22 mRNA by ABA was inhibited by cycloheximide. Thus, it appears that protein synthesis is required for the induction of this mRNA by ABA. By contrast, protein synthesis was not required for the ABA-responsive induction RD29 mRNA, which corresponds to another dehydration-responsive gene of A. thaliana. These results suggest that there are at least two mechanisms for the induction of dehydration-responsive genes by ABA. RD22 mRNA was also expressed during the early and middle stages of seed development, showing a pattern of expression similar to that of USP. The seed-specific expression of RD22 seems not to be regulated by ABA. Structural analysis of the RD22 genomic clone revealed that the structural gene (designated rd22) contains three introns, and only a single copy of the gene is present in the A. thaliana genome, while the gene for USP from V. faba is actually a family of genes with 10 to 20 members.(ABSTRACT TRUNCATED AT 250 WORDS)
Mol Gen Genet 1993 Apr
PMID:The plant hormone abscisic acid mediates the drought-induced expression but not the seed-specific expression of rd22, a gene responsive to dehydration stress in Arabidopsis thaliana. 847 24

Cells of a heat-shock resistant mutant were approximately 1000-times more resistant to lethal heat shock than those of the parental strain. We observed that exponentially growing cells of the mutant synthesized trehalose and showed increased osmotolerance, dehydration tolerance an ethanol tolerance, a fact not observed in wild type strains. The mutant synthesizes constitutively six proteins, among them two proteins of 56 and 63 kDa. Interestingly these molecular weights could correspond to the subunit of trehalose-6-phosphate synthase and to phosphoglucomutase II, respectively. Our results showed that glucose-growing cells of the hsr 1 mutant possessed high levels of activity of these enzymes when compared to the control strain.
Biochem Mol Biol Int 1995 Aug
PMID:Effect of trehalose during stress in a heat-shock resistant mutant of Saccharomyces cerevisiae. 853 93

Fusion of African swine fever virus (ASFV) with model membranes was monitored by a fluorescence dequenching assay. ASFV was able to fuse with liposomes of various compositions. Fusion was more extensive with liposomes made of negatively charged phospholipids, and reduced in the presence of the neutral phospholipid phosphatidylcholine (PC). Fusion activity of ASFV was pH-dependent, the extent of fusion increasing with decreasing pH for all target membranes. These results are consistent with the in vivo characteristics of fusion of ASFV with the endosome membrane. An increase in fusion activity was obtained with dehydrating agents, similarly to what occurs with other lipid-enveloped viruses. Dehydrating agents such as dimethylsulfoxide and dimethylsulfone greatly enhanced fusion, the effect of dimethylsulfone being more pronounced than that of dimethylsulfoxide. Poly(ethylene glycol) also potentiated ASFV fusion activity, and the effect of this polymer was found to be dependent on its molecular weight. These results stress the importance of dehydration and hydrophobic interactions on the early events of viral penetration into target cells.
Biochem Mol Biol Int 1995 Oct
PMID:Fusion activity of African swine fever virus towards target membranes: lipid dependence and effect of dehydrating agents. 859 88

Trg-31 is a pea gene which is rapidly induced in response to dehydration stress. Trg-31 belongs to a family of membrane intrinsic proteins that play a role in facilitating inter-membrane transport. In order to understand the mechanism of regulation of Trg-31 expression we defined the cis-acting elements of the Trg-31 promoter and the trans-acting factors that interact with them. Deletion analysis of the Trg-31 promoter in transgenic tobacco plants indicated the presence of two positive regulatory region: (1) between the -1412 to -1066 nucleotides and (2) between the -559 to -391 nucleotides, with respect to the transcription initiation site. A negative regulatory element was also identified between -1066 and -559 nucleotides. All three regulatory regions specifically regulated Trg-31 induction in leaf tissue and had little or no effect on Trg-31 induction in root tissue. Gel-mobility shift assays indicated the presence of leaf nuclear proteins that interacted with the -1412 to -1066 and -1066 to -559 regions and an 'ABRE-like' sequence at -74 in the 5' promoter region of Trg-31.
Plant Mol Biol 1996 Mar
PMID:Identification of tissue-specific, dehydration-responsive elements in the Trg-31 promoter. 870 33

Musca domestica trypsin antibody cross-reacts with polypeptide bands of M(r) 25,000 and 30,000 showing proteolytic activity from Stomoxys calcitrans midgut extracts. Secretory granules from the main enzyme-secreting region, the opaque zone, stained heavily with the trypsin antibody in both unfed and blood-fed flies. Heterogeneous staining of granules suggests the unequal distribution of trypsin in secretory granules. This is also consistent with the occurrence of non-parallel secretion, which is also suggested by the possible preferential release of smaller, heavily stained secretory granules in fed flies. The predigestive, anterior midgut region responsible for rapid dehydration of the blood meal, the reservoir zone, contains a different population of secretory granules which stain heavily with trypsin antibody. This zone contains 20% of the midgut trypsin activity in unfed flies; trypsins are held here as proenzymes which are probably only activated postsecretion. In the midgut lumen of both unfed and blood-fed flies, trypsin is mainly immunolocalized in the ectoperitrophic space. Enzyme assays suggest that 5-15% of the lumenal trypsin is associated with the peritrophic matrix. The finding of intact secretory granules plus cell debris in the ectoperitrophic space of opaque and lipoid zones of blood-fed flies supports the contention that some trypsin is released by apocrine secretion in this insect.
Insect Biochem Mol Biol 1996 May
PMID:An immunocytochemical investigation of trypsin secretion in the midgut of the stablefly, Stomoxys calcitrans. 876 63

We have isolated a gene and cDNA from Brassica napus encoding a hybrid-proline-rich protein. The putative protein is modular in structure. The N-terminal domain has properties of a signal peptide which would direct the protein into the ER. Amino acids 27 to 287 comprise three domains which contain high levels of proline and several other amino acids common in proline-rich cell wall proteins. These domains are characterised by repeating amino acid motifs. The C-terminal domain (amino acids 288 to 376) contains three putative membrane-spanning regions and shows a high degree of amino acid similarity to known hybrid-proline-rich proteins from several species. It is likely that the protein is secreted from the cell, located in the cell wall and anchored in the plasma membrane via the C-terminal domain. Transcripts encoding this protein are induced in leaf tissue within 8 h of cold treatment and decrease rapidly when plants are returned to normal temperatures. The transcripts are not induced by heat shock, dehydration, exogenous ABA or wounding, whereas transcripts of a control B. napus gene are induced by dehydration and ABA. The possible function of this protein in cold tolerance is discussed.
Plant Mol Biol 1996 Jul
PMID:Transcripts of a gene encoding a putative cell wall-plasma membrane linker protein are specifically cold-induced in Brassica napus. 880 8

Coding sequences for two cysteine proteases were amplified from cDNA derived from pea nodule mRNA using primers based on conserved regions of plant cysteine proteases. One of the amplified cDNA sequences corresponded to a previously described cysteine protease gene, Cyp15a, expressed in pea shoots in response to dehydration (J.T. Jones and J.E. Mullet, Plant Mol. Biol. 28:1055-1065, 1995). Inside the pea root nodule, in situ hybridization revealed that this gene is expressed strongly in the apical region and more weakly in the uninfected cortex and in the central infected tissue where nitrogen fixation takes place. The complete sequence of the cDNA corresponding to the other gene, PsCyp1, was obtained. Expression of this gene, which was studied both on RNA blots and in situ, showed good correlation with the onset of nodule senescence. In situ hybridization studies revealed that PsCyp1 was expressed in senescent infected tissue at the base of the nodule. This signal was just detectable in normal symbiotically wild-type nodules but was much stronger in the early senescing nodules formed by a symbiotically defective mutant of Rhizobium leguminosarum.
Mol Plant Microbe Interact 1996 Nov
PMID:Expression of cysteine protease genes in pea nodule development and senescence. 887 Feb 68


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