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Query: UNIPROT:P06889 (Mol)
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Using immunohistochemical techniques, the keratin expression patterns in basal and columnar cells (mucus-producing and ciliated cells) were investigated in tracheal organ cultures. Tracheas were from either hamsters fed a control diet or from hamsters fed a vitamin A-deficient diet; tracheas from the latter group were treated in vitro with all-trans retinol. In tracheas from hamsters fed a control diet, basal cells generally reacted with the RCK102 antibody and columnar cells with the RGE53 and the HCK19 antibodies, and both basal and columnar cells were recognized by the RCK105 antibody. The squamous cell cytokeratin 10 (detected by the RKSE60 antibody) was not expressed in cultured tracheas from hamsters fed a normal or a vitamin A-deficient diet. In the course of the in vitro period a number of keratins were "switched on" or "switched off" in both basal and columnar cells. In tracheas from vitamin A-deprived hamsters the RCK102 antibody clearly recognized basal cells and cigarette smoke condensate-induced proliferating basal cells, whereas the RGE53 antibody reacted with mucus-producing and ciliated cells. During organ culture foci of columnar epithelial cells expressed basal cell properties (detected with the RCK102 antibody) after all-trans retinol treatment and were found negative for the RGE53 antibody. Furthermore, it appeared that the RGE53-negative columnar cells contained periodic acid-Schiff-positive mucous granules. These findings indicate that basal cells may differentiate into columnar cells. Tracheal epithelium did not appear to co-express vimentin next to keratins during organ culture, which may be due to the intact three-dimensional organization present in these organ cultures.
Virchows Arch B Cell Pathol Incl Mol Pathol 1988
PMID:Intermediate filament expression in normal and vitamin A depleted cultured hamster tracheal epithelium as detected by monoclonal antibodies. A study with emphasis on histological changes. 246 33

The effects of cigarette smoke condensate (CSC) and all-trans retinol on the cell proliferative activity of vitamin A-deprived hamster tracheal epithelium have been studied in vitamin A-deficient, serum-free, hormone-supplemented medium in organ culture. In the absence of retinol, CSC induced a dose-dependent increase in labeling index (LI) during 12 days of culture. The basal cells were more sensitive to CSC exposure than non-basal cells during the first 6 to 8 culture days. However, in squamous metaplastic foci developing after culture day 6, both basal and non-basal cells in the mid-part of the epithelium were labeled. Physiological concentrations of all-trans retinol stimulated the non-basal LI and inhibited the basal cell LI. Compared with dimethylsulfoxide (DMSO), all retinol concentrations used in the present study inhibited the basal cell LI at each time point examined (4-12 days culture). Exposure of tracheal rings to retinol, either before or after exposure to CSC, or simultaneous exposure to retinol and CSC, clearly decreased the CSC-induced basal cell proliferative activity depending on the retinol concentration used. It is concluded from the present study that squamous metaplasia induced by vitamin A-deficiency or by CSC originates mainly from basal cells and that for the maintenance of these lesions, both basal and non-basal cells play a role. Furthermore, all-trans retinol inhibited CSC-induced basal cell proliferation.
Virchows Arch B Cell Pathol Incl Mol Pathol 1988
PMID:Effects of all-trans retinol and cigarette smoke condensate on hamster tracheal epithelium in organ culture. I. A cell proliferation study. 289 24

The effects of all-trans retinol and cigarette smoke condensate (CSC) on tissue morphology and cellular differentiation were investigated in vitamin A-deprived tracheal epithelium cultured in vitamin A-and serum-free hormone-supplemented medium. Physiological retinol concentrations prevented the development of hyperplasia and squamous metaplasia with or without keratinization, and induced differentiation to mucous cells. Squamous metaplastic foci with keratinization were observed during 12 days of culture with low retinol concentrations and with dimethylsulfoxide (DMSO) which was accompanied by an increased number of basal and indeterminate cells. CSC induced a dose-related hyperplasia and irregularly shaped foci of squamous metaplasia with atypical epithelial proliferation. In non-metaplastic epithelium, CSC exposure increased the number of ciliated cells. Hyperplasia and squamous metaplasia were inhibited if the tracheal rings were first treated with retinol followed by CSC exposure, or if the tracheas were simultaneously treated with retinol and CSC. CSC-exposure prior to retinol treatment induced similar histomorphological alterations as CSC alone.
Virchows Arch B Cell Pathol Incl Mol Pathol 1988
PMID:Effects of all-trans retinol and cigarette smoke condensate on hamster tracheal epithelium in organ culture. II. A histomorphological study. 289 25

Possible steps in the folding of bacteriorhodopsin are revealed by studying the refolding and interaction of two fragments of the molecule reconstituted in lipid vesicles. (1) Two denatured bacteriorhodopsin fragments have been purified starting from chymotryptically cleaved bacteriorhodopsin. Cleaved bacteriorhodopsin has been renatured from a mixture of the fragments in Halobacterium lipids/retinal/dodecyl sulfate solution following removal of dodecyl sulfate by precipitation with potassium. The renatured molecules have the same absorption spectrum and extinction coefficient as native cleaved bacteriorhodopsin. They are integrated into small lipid vesicles as a mixture of monomers and aggregates. Extended lattices form during the partial dehydration process used to orient samples for X-ray and neutron crystallography. (2) Correct refolding of cleaved bacterioopsin occurs upon renaturation in the absence of retinal. Regeneration of the chromophore and reformation of the purple membrane lattice are observed following subsequent addition of all-trans retinal. (3) The two chymotryptic fragments have been reinserted separately into lipid vesicles and refolded in the absence of retinal. Circular dichroism spectra of the polypeptide backbone transitions indicate that they have regained a highly alpha-helical structure. The kinetics of chromophore regeneration following reassociation have been studied by absorption spectroscopy. Upon vesicle fusion, the refolded fragments first reassociate, then bind retinal and finally regenerate cleaved bacteriorhodopsin. The complex formed in the absence of retinal is kinetically indistinguishable from cleaved bacterioopsin. The refolded fragments in lipid vesicles are stable for months, both as separate entities and after reassociation. These observations provide further evidence that the native folded structure of bacteriorhodopsin lies at a free energy minimum. They are interpreted in terms of a two-stage folding mechanism for membrane proteins in which stable transmembrane helices are first formed. They subsequently pack without major rearrangement to produce the tertiary structure.
J Mol Biol 1987 Dec 20
PMID:Refolding of bacteriorhodopsin in lipid bilayers. A thermodynamically controlled two-stage process. 343 Jun 24

The dark-adapted form of bacteriorhodopsin in the purple membrane of Halobacterium halobium changes its absorption maximum from 560 to 600 nm if the pH is lowered to about 2 [Oesterhelt, D., & Stoeckenius, W. (1971) Nature (London), New Biol. 233, 149; Moore, T. A., Edgerton, M. E., Parr, G., Greenwood, C., & Perham, R. N. (1978) Biochem. J. 171, 469; Mowery, P. C., Lozier, R. H., Chae, Q., Tseng, T.-W., Taylor, M., & Stoeckenius, W. (1979) Biochemistry 18, 4100; Fischer, U., & Oesterhelt, D. (1979) Biophys. J. 28, 211; Muccio, D. D., & Cassim, J. Y. (1979) J. Mol. Biol. 135, 595]. We compared the pH dependence of the absorption spectra of acetylated membrane with that of unacetylated native membrane. The completely acetylated membrane showed a midpoint of pH 4.8 for the conversion to the acidic form; that of the native membrane was 3.4. On acetylation, the absorption maximum at neutral pH moved from 560 to 555 nm with about 20% decreases in extinction coefficients as compared with that of the native membrane, whereas the spectrum in acid was not affected. The chloride-dependent blue shift from the acidic form of the acetylated membrane was largely suppressed. The CD spectrum of the acetylated membrane was composed of two bands of an opposite sign with slightly decreased amplitudes. The chromophore of the acetylated membrane was sensitive to hydroxylamine, and the spectrum before bleaching was restored on addition of all-trans-retinal to the bleached membrane followed by dark incubation. Blue light irradiation accelerated the conversion to the acidic form in the native membrane but not in the acetylated membrane. Reductive ethylation did not affect the pH dependence of the absorption spectra.
...
PMID:Absorption spectral properties of acetylated bacteriorhodopsin in purple membrane depending on pH. 712 52

It was shown that photoregeneration of rhodopsin from metarhodopsin II takes place in bovine retinal rod outer segment disc membrane fragments. Photoregenerated rhodopsin is identical to unbleached rhodopsin in spectra and in stability to NH2OH. Moreover, as an unbleached pigment, photoregenerated rhodopsin can induce the transient increase of the artificial lipid membrane (ALM) conductivity in response to visible light. It was shown, that the UV-light converts metarhodopsin II to rhodopsin through the new long-lived (tau approximately 5 min) intermediate product (X500). X500 is indistinguishable from rhodopsin spectrophotometrically, but it can not induce the transient increase of ALM conductivity in response to visible light. It was shown that P467 (this product is obtained usually by photolysis of metarhodopsin II) has a retinal chromophore in all-trans configuration. This result indicates that P467 is produced from cis-metarhodopsin II and X500 (these products are the intermediates of the back reaction metarhodopsin II--rhodopsin) but not from metarhodopsin II as suggested earlier. A modified scheme of the back reaction metarhodopsin II--rhodopsin is given.
Mol Biol (Mosk)
PMID:[Molecular mechanisms of photoreception. IV. Photoregeneration of rhodopsin from metarhodopsin II using the artificial lipid membrane method for detection of intermediate steps of this reaction]. 732 16

All-trans-3,7,11,15-tetramethyl-2,4,6,10,14-hexadecapentaenoic acid (designated "acyclic retinoid") induced upregulation of the albumin gene expression at its transcriptional level, whereas all-trans-retinoic acid (RA) induced downregulation of the expression in both PLC/PRF/5 and HuH7 human hepatoma cell lines. These up- and down regulations of the albumin gene expression coordinated with high and low levels of mRNA for hepatocyte nuclear factor-1 (HNF-1), which is one of the most potent transcription factors for the albumin gene, implying that retinoids may regulate albumin gene expression through HNF-1 expression in opposite ways. The PLC/PRF/5 and HuH7 hepatoma cell lines expressed retinoid X receptor-alpha (RXR alpha) mRNA, whose expression was constitutive. Acyclic retinoid and all-trans-RA both induced upregulation of retinoic acid receptor-beta (RAR beta), and both suppressed cell proliferation-related phenotypic expressions by the alpha-fetoprotein gene and the c-myc oncogene. 9-cis-RA, whose receptor is known to be RXR alpha, also induced upregulation of albumin and HNF-1 expression. These results suggest that acyclic retinoid may act through both RXR alpha and RAR beta, whereas all-trans-RA conveys only RAR beta-mediated functions, at least in these two hepatoma cell lines.
Mol Carcinog 1994 Jul
PMID:Positive and negative regulations of albumin gene expression by retinoids in human hepatoma cell lines. 751 16

The human acute promyelocytic leukemia (APL) cell line HL-60 differentiates to functionally mature granulocytes by incubation with all-trans-retinoic acid (RA). Since T3 and RA are important in cell differentiation and development, and since their receptors are highly homological, we investigated the T3 effects on RA-induced HL-60 cell differentiation. Although T3 alone did not induce cell differentiation, RA-mediated differentiation was significantly enhanced in the presence of 10(-7) M T3. This effect of T3 was considered to be mediated, at least in part, by increased intracellular cAMP, since the phosphodiesterase inhibitor enhanced, and the protein kinase A antagonist partially blocked, T3 potentiation. When HL-60 cells were pretreated with RA for 20 h, T3 alone stimulated the cell differentiation. The time-course study showed that incubation with RA for 12 h was necessary for HL-60 cells to be primed to respond to T3 for differentiation. The present finding that T3 potentiates RA-induced HL-60 cell differentiation may raise the possibility that T3 supplement increases clinical remission in APL patients who are treated with RA.
Mol Cell Endocrinol 1994 Jul
PMID:3,5,3'-Triiodothyronine stimulates retinoic acid-induced differentiation in HL-60 cells. 752 82

In McA-RH 8994 rat hepatoma cells, all-trans-retinoic acid (t-RA) induces expression of the alpha-fetoprotein (AFP) and albumin genes and results in a phenotype similar to differentiated fetal hepatocytes. The present study elucidated the mechanism involved in AFP gene regulation mediated by retinoic acid. Northern blot analyses demonstrated that 9-cis-retinoic acid (c-RA), a ligand for retinoid x receptors (RXRs), also induced expression of the AFP gene in McA-RH 8994 cells. The induction was time- and dose-dependent. Northern blots and transfection assays using the 7.3 kb full-length regulatory region of the AFP gene demonstrated that c-RA was more effective than t-RA in regulating expression of the AFP gene. At 10(-7) M, c-RA increased AFP mRNA 5-fold and chloramphenicol acetyltransferase (CAT) activity 2.5-fold. In contrast, t-RA at a concentration of 10(-7) M exerted no significant effect; 10(-6) to 10(-5) M t-RA was needed to affect AFP gene expression. These data suggested that activation of RXRs is essential for the regulation of the AFP gene. Co-transfection experiments revealed that over-expression of RXR alpha in McA-RH 8994 cells further enhanced the CAT activity induced by c-RA. In addition, c-RA did not alter the half-life of AFP mRNA. Thus, RXR alpha may play a crucial role in transcriptional regulation of the AFP gene and in controlling hepatocyte phenotype.
J Mol Endocrinol 1995 Feb
PMID:9-cis-retinoic acid is more effective than all-trans-retinoic acid in upregulating expression of the alpha-fetoprotein gene. 753 13

A recombinant form of murine apo-cellular retinoic acid binding protein I (apo-CRABPI) has been purified and crystallized at pH 5.0, and the crystal structure has been refined to an R-factor of 19.6% at a resolution of 2.7 A. CRABPI binds all-trans retinoic acid and some retinoic acid metabolites with nanomolar affinities. Coordinates of the holo form of CRABP were not available during the early stages of the study, and in spite of numerous homologs of known structure, phases were not obtainable through molecular replacement. Instead, an interpretable electron density map was obtained by multiple isomorphous replacement methods after improvement of the heavy-atom parameters with density modified trial phases. Two molecules of apo-CRABPI occupy the P3121 asymmetric unit and are related by pseudo 2-fold rotational symmetry. Unique conformational differences are apparent between the two molecules. In all of the family members studied to date, there is a lack of hydrogen bonds between two of the component beta-strands resulting in a gap in the interstand hydrogen bonding pattern. In the crystallographic dimer described here, a continuous intermolecular beta-sheet is formed by using this gap region. This is possible because of an 8 A outward maximum displacement of the tight turn between the third and fourth beta-strands on one of the molecules. The result is a double beta-barrel containing two apo-CRABPI molecules with a more open, ligand-accessible binding cavity, which has not been observed in other structures of a family of proteins that bind hydrophobic ligands.
J Mol Biol 1995 Sep 29
PMID:Crystal structure of cellular retinoic acid binding protein I shows increased access to the binding cavity due to formation of an intermolecular beta-sheet. 756 63


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