RESUMEN
This article begins by presenting corpus linguistics and explaining how the corpus approach to language analysis can be used to investigate the language of dentistry. It then reports findings from an analysis that was carried out with readily available software and a corpus of 1,400 dental research abstracts. Included is information about word frequency in dental abstracts and also about how certain words tend to co-occur in sentences and form regular patterns. The article concludes with a discussion of ways that corpus-based findings can be applied to the teaching of English to non-native speakers of the language, many of whom will need English skills for their future careers in dentistry.
Asunto(s)
Odontología , Lenguaje , Lingüística/métodos , Indización y Redacción de Resúmenes , Educación en Odontología , Lingüística/estadística & datos numéricos , América del Norte , Publicaciones Periódicas como Asunto , Programas Informáticos , Terminología como AsuntoRESUMEN
The small, proline-rich (SPR) genes consist of three subclasses closely linked on human chromosome 1, a region referred to as the epidermal differentiation complex. SPR genes consist of two exons, with the second exon containing the entire open reading frame. SPRs are expressed in all squamous tissues of the skin, scalp, footpad, vaginal epithelia, and most of the epithelial lining of the digestive tract, including the lip, tongue, esophagus, and forestomach. Although SPR1 is absent in normal mucociliary epithelium of the respiratory tract, epithelia that undergo squamous differentiation in response to vitamin-A deficiency or to injury owing to exposure to environmental toxicants express SPR1. High levels of SPR1 are detected in various diseases and cancers of the skin or respiratory epithelia and in nonkeratinizing papillary adenocarcinomas. SPR expression can be regulated by transcriptional factors, by posttranscriptional factors, or by factors that affect SPR1 mRNA translation or protein turnover. Furthermore, regulation can be affected by the state of cell proliferation. The presence of SPR1 in most of these epithelia, and the absence of SPR3 in normal skin, suggest that these subclasses have distinct functions. Various approaches to the study of the cross-linked envelope (CE) components in identifying SPR1 and SPR2 and in suggesting that SPRs are one of the precursor proteins of the CE. However, expression of SPR1 in nonsquamous tissues and cell lines indicates a function not associated with squamous differentiation. Several studies have demonstrated that SPR1 antibodies react with nuclear proteins and that SPR1 is expressed in cells before entering the G0 phase of the cell cycle. Future studies should clarify the role of SPRs by modifying their contents in CE, and should identify SPR-associated proteins to clarify the cell growth-related role of SPR1.
Asunto(s)
Regulación Neoplásica de la Expresión Génica , Proteínas de Filamentos Intermediarios/metabolismo , Familia de Multigenes , Proteínas/metabolismo , Envejecimiento , Secuencia de Aminoácidos , Animales , Células CHO , Calcio/metabolismo , Recuento de Células , División Celular , Secuencia de Consenso , Proteínas Ricas en Prolina del Estrato Córneo , Cricetinae , Citocinas/metabolismo , Sistema Digestivo/metabolismo , Epidermis/metabolismo , Humanos , Neoplasias Pulmonares/patología , Proteínas de la Membrana , Datos de Secuencia Molecular , Neoplasias de Células Escamosas/patología , Sistema Respiratorio/metabolismo , Retinoides/metabolismo , Homología de Secuencia de Aminoácido , Piel/metabolismo , Neoplasias Cutáneas/metabolismo , Acetato de Tetradecanoilforbol/metabolismo , Factores de Tiempo , Células Tumorales CultivadasRESUMEN
Dramatic morphological, biochemical and cytological changes occur in parotid glands of rats and mice which have been treated with the beta-adrenergic receptor agonist isoproterenol (Ipr). Changes include glandular hypertrophy, induction of tissue-specific proline-rich proteins (PRPs), increases in cAMP, and occurrence of polyploidy. Similar changes also occur after feeding mice polyphenolic compounds commonly referred to as tannins. Data are presented which show that changes in cell cycle proteins are due to stimulation of the beta-adrenergic receptor by either isoproterenol or tannin treatment of mice. Both p34cdc2 mRNA and protein levels were elevated dramatically after mice were treated. Induction of p34cdc2 occurred within 24 hrs. and was transient during treatment. The beta1-adrenergic receptor antagonist atenolol blocked both tissue-specific expression of proline-rich proteins and induction of p34cdc2. Coincident with the increase in p34cdc2, cyclin-dependent kinase complexes containing cyclins A and B increased forty- and ten-fold, respectively. These results show that in mouse parotid glands activation of the beta1-adrenergic receptor by either the administration of isoproterenol or ingestion of dietary tannins induces synthesis of p34cdc2 and most likely contributes to the occurrence of polyploidy.
Asunto(s)
Agonistas Adrenérgicos beta/farmacología , Proteína Quinasa CDC2/biosíntesis , Isoproterenol/farmacología , Glándula Parótida/efectos de los fármacos , Receptores Adrenérgicos beta 1/efectos de los fármacos , Proteínas y Péptidos Salivales/biosíntesis , Transducción de Señal/efectos de los fármacos , Antagonistas Adrenérgicos beta/farmacología , Animales , Areca/efectos adversos , Atenolol/farmacología , Proteína Quinasa CDC2/genética , Ciclo Celular/efectos de los fármacos , Cromatografía de Afinidad , Ciclina A/metabolismo , Ciclina E/metabolismo , Inducción Enzimática/efectos de los fármacos , Taninos Hidrolizables/farmacología , Ratones , Ratones Endogámicos A , Glándula Parótida/enzimología , Plantas Medicinales , Receptores Adrenérgicos beta 1/fisiologíaRESUMEN
Proline-rich protein mRNAs are increased dramatically in the salivary glands of rats, mice, and hamsters upon treatment with the beta-agonist isoproterenol. Sequence comparisons between mice and hamster proline-rich protein genes identified conserved regions upstream from the transcription start site. Reporter plasmids containing these 5'-flanking sequences from a mouse proline-rich protein gene, MP2, were constructed and tested for transcriptional regulation by cAMP. Transient transfection experiments in mouse L-M cells showed that the upstream region -702 to -322 bp relative to the transcription start site is sufficient to confer cAMP induction on a heterologous promoter. Multiple copies of the AP-1 sequence elements within this region (-625 to -551) mediate the cAMP transcriptional response of reporter gene expression in L-M cells. L-M cell nuclear proteins and purified human c-jun protein bind to these upstream elements as determined by DNase I footprint analysis. Nuclear proteins isolated from mouse parotid glands protected the consensus AP-1 binding site 5'-TGAGTCA-3' (-592 to -586). The nuclear proteins interacting at this site were increased about sixfold in glands isolated from isoproterenol-treated mice when compared with glands from untreated mice. These results suggest that induction of AP-1 transcription factors in the parotid gland control the upregulation of some mouse salivary proline-rich proteins.
Asunto(s)
AMP Cíclico/metabolismo , Glándula Parótida/metabolismo , Péptidos/genética , Factor de Transcripción AP-1/biosíntesis , Agonistas Adrenérgicos beta/farmacología , Animales , Secuencia de Bases , Sitios de Unión/genética , Cloranfenicol O-Acetiltransferasa/genética , Secuencia de Consenso , Cricetinae , ADN/genética , ADN/metabolismo , Sondas de ADN/genética , Regulación de la Expresión Génica , Genes Reporteros , Humanos , Isoproterenol/farmacología , Ratones , Glándula Parótida/efectos de los fármacos , Dominios Proteicos Ricos en Prolina , Regiones Promotoras Genéticas , ARN Mensajero/genética , ARN Mensajero/metabolismo , RatasRESUMEN
A family of small proline-rich proteins (SPR1s) is induced in cells undergoing squamous differentiation. Because SPR1 mRNA is detected in mesenchymal nasal cells of rats exposed to cigarette smoke, expression of this mRNA in other nonsquamous cells and tissues was investigated. Using PCR, low levels of SPR1 mRNA were identified in a number of nondifferentiating cell lines and in nonsquamous tissues. G(0)SPR1 mRNA, the hamster homologue of SPR1 mRNA, was increased 10-fold in Chinese hamster ovary (CHO) cells when the culture reached 80-90% confluence and was downregulated after cells ceased growing at 100% confluence. The deduced amino acid sequence of G(0)SPR1 showed a high homology to the family of SPR1 from different species. Affinity-purified antibodies to SPR1 reacted to about 50% of the CHO cell population, indicating that the protein is expressed at specific stages of the cell cycle. CHO cells that were switched to low-serum medium when they were at 60% confluence showed an increase in G(0)SPR1 levels before the cells entered G0, indicating that G(0)SPR1 may b a signal to cells entering G0. Because expression of the SPR1 family of proteins is associated with squamous differentiation, the observations in the nondifferentiating CHO cells indicate that these proteins may play a role in mediating the withdrawal from the cell cycle prior to the commitment to differentiation.
Asunto(s)
Ciclo Celular , Biosíntesis de Proteínas , Transcripción Genética , Secuencia de Aminoácidos , Animales , Anticuerpos , Secuencia de Bases , Células CHO , División Celular , Proteínas Ricas en Prolina del Estrato Córneo , Cricetinae , Cartilla de ADN , Fase G1 , Haplorrinos , Humanos , Immunoblotting , Cinética , Proteínas de la Membrana , Datos de Secuencia Molecular , Fragmentos de Péptidos/química , Fragmentos de Péptidos/inmunología , Reacción en Cadena de la Polimerasa , Proteínas/análisis , Proteínas/química , ARN Mensajero/biosíntesis , Conejos , Ratas , Fase de Descanso del Ciclo Celular , Homología de Secuencia de Aminoácido , PorcinosRESUMEN
The absence of vitamin A or vitamin A derivatives in culture media promotes squamous cell differentiation of tracheobronchial epithelial cells. This is especially true for the expression of a small proline-rich protein (20K; 98 amino acids) in pig trachea epithelial cells. Multigene families encode different small proline-rich proteins in different species, and these proteins are possible markers for squamous cell differentiation. 20K mRNA and 20K protein were detected in cells within 4 and 5 days in culture, respectively, when cells reached about 50% confluence, and expression increase 12-fold during cell proliferation until cells reached 100% confluence. Arotinoid (10(-9)M), a synthetic retinoid, essentially totally inhibited expression of 20K mRNA in proliferating tracheobronchial cells within 3 days of treatment while 20K protein levels were only decreased 4-fold after 5 days. However, if cells were exposed to arotinoid 3 days after reaching confluent growth, the levels of either 20K mRNA or 20K protein were unchanged. Cells exposed to arotinoid from the onset of culturing, and then removal of the retinoid from proliferating cells resulted in the expression of 20K mRNA and protein after 4 and 5 days as observed previously. 20K mRNA was not detected in cells that had been continuously exposed to arotinoid from the start of culture until 3 days post confluence, even 10 days following removal of arotinoid. Our results strongly suggest that the growth phase and state of cell differentiation greatly affect the response of these epithelial cells to vitamin A derivatives.
Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Proteínas/genética , Retinoides/farmacología , Tráquea/metabolismo , Secuencia de Aminoácidos , Animales , División Celular , Células Cultivadas , Proteínas Ricas en Prolina del Estrato Córneo , Células Epiteliales , Epitelio/metabolismo , Proteínas de la Membrana , Datos de Secuencia Molecular , Biosíntesis de Proteínas , ARN Mensajero/biosíntesis , Porcinos , Tráquea/citologíaRESUMEN
In cell-free translations of RNA from primary cultures of pig trachea surface epithelial cells we observed that a mRNA encoding a 20 kDa proline-rich protein (sPRP) was dramatically induced during culturing (Tesfaigzi et al., 1990, Biochem. Biophys. Res. Commun., 172:M1304-1309). This mRNA was not detected in tracheal tissue or in epithelial cells prior to culturing. Antisera were raised to synthetic peptide sequences corresponding to 23 amino acids on the C-terminus (C23-antiserum) and 29 amino acids on the N-terminus (N29 antiserum) of sPRP. On Western blot analysis, C23 antiserum reacted with a 20 kDa protein in cytosolic extracts from pig tracheal cells maintained in culture for 4 days. The reaction with the 20 kDa protein was inhibited by adding C23 peptide. Two nuclear proteins (66 and 70 kDa) obtained by micrococcal nuclease treatment of tracheal cell nuclei were detected on Western blots with C23 antiserum. These proteins were present in cells both before and after culturing. Sucrose gradient fractionation indicated that these nuclear proteins are associated with chromatin. Small amounts of the 66 and 70 kDa proteins were obtained from nuclear matrix fractions. These nuclear proteins also reacted with N29 antiserum. Since these proteins share similar epitopes with the N- and C-termini of sPRP, it is likely that the 20 kDa protein (sPRP) is part of these proteins. However, purification of the nuclear proteins followed by an amino acid sequence analysis is necessary to clarify whether sPRP is part of these proteins.
Asunto(s)
Diferenciación Celular , Proteínas Nucleares/inmunología , Péptidos/inmunología , Tráquea/inmunología , Secuencia de Aminoácidos , Animales , Citosol/química , Epitelio/inmunología , Técnica del Anticuerpo Fluorescente , Datos de Secuencia Molecular , Peso Molecular , Matriz Nuclear/química , Péptidos/química , Péptidos/metabolismo , Dominios Proteicos Ricos en Prolina , PorcinosRESUMEN
Eight subjects (5 men, 3 women, ages 27 to 55) with obstructive sleep apnea syndrome (OSAS) were studied to quantify and compare electromyographic (EMG) activity of levator veli palatini (LVP) and palatoglossus (PG), two velopharyngeal muscles, and genioglossus (GG) during obstructive apnea cycles in non-rapid eye movement (NREM) sleep. EMG activity of three successive preapneic breaths, first and last apneic efforts, and three successive postapneic breaths was quantified for each muscle as peak phasic inspiratory EMG normalized as percent activity of the last preapneic breath. In all subjects, apnea onset coincided with simultaneous inspiratory EMG nadir of all three muscles (LVP = 63 +/- 40%, PG = 74 +/- 53%. GG = 83 +/- 48%. mean +/- SD activity of last preapneic breath). Apnea resolution did not occur until inspiratory EMG of all three muscles simultaneously reached maximal activity, at levels significantly greater than preapneic activity as well as activity of the last preapneic effort (LVP = 215 +/- 205%, PG = 227 +/- 240+, GG = 235 +/- 202%, mean +/- SD activity of last preapneic breath, p < 0.05, Fisher's partial least-squares difference [PLSD] test for each muscle). The presence or absence of electroencephalographic arousal at apnea resolution did not influence these patterns of EMG activity. Inspiratory recruitment of velopharyngeal as well as oropharyngeal muscles appears to be associated with upper airway patency during sleep in patients with OSAS.
Asunto(s)
Músculos Palatinos/fisiopatología , Respiración/fisiología , Síndromes de la Apnea del Sueño/fisiopatología , Adulto , Electromiografía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Paladar Blando/fisiología , PolisomnografíaRESUMEN
Sturgeon are an ancient family (Acipenseridae) of fishes that lie close to the divergence of fish that eventually evolved into terrestrial animals and those that evolved into modern teleost species. Therefore, white sturgeon vitellogenin sequences fill a gap in the current understanding of the functional domains of this protein family. Vitellogenin cDNA was sequenced and used to investigate gene expression in white sturgeon, Acipenser transmontanus. Estrogen-induced vitellogenin mRNA was detected in the livers of both males and females and was also detected in undifferentiated gonads of estrogen-treated fish. Low levels of vitellogenin mRNA were also detected in the testis of both control and estrogen-treated males. The cDNA encoded a 186-kDa protein that was missing only six to seven of the amino-terminal amino acids. Comparisons to silver lamprey, Xenopus, and chicken vitellogenin sequences indicate that the overall structure of the yolk protein domains were highly conserved. There was considerable homology in three regions of the lipovitellin I domain. These conserved sequences are likely to be involved in vitellogenin receptor binding. The phosvitin domain of white sturgeon vitellogenin contained fewer and shorter serine repeats as predicted from yolk protein phosphate content of fish compared to Xenopus and chicken. However, the vitellogenin of white sturgeon had a lower serine content as compared with silver lamprey, indicating that an increased serine content is not strictly a function of evolutionary age.
Asunto(s)
Aminoácidos/análisis , Peces/genética , Hígado/metabolismo , Vitelogeninas/biosíntesis , Vitelogeninas/genética , Secuencia de Aminoácidos , Animales , Pollos/genética , ADN Complementario , Femenino , Expresión Génica , Lampreas/genética , Masculino , Datos de Secuencia Molecular , ARN Mensajero/biosíntesis , Homología de Secuencia de Aminoácido , Caracteres Sexuales , Xenopus/genéticaRESUMEN
The cDNAs for two glycoproteins, the 158-kDa submandibular glycoprotein (SGP158) and the 200-kDa parotid glycoprotein (PGP200), have been cloned from rat submandibular and parotid glands, respectively. Both cDNAs encode for identical proteins with repeating peptides -Asp-Gln-Gly-(Asn)-Gln-Thr-Gln-Pro-Arg-Pro-Pro-His-Pro-. A full-length cDNA encoding SGP158 was obtained using the strategy of anchor-PCR, and a full-length cDNA of PGP200 was prepared using RNA-PCR. Sequence analysis of the cDNAs revealed that SGP158 and PGP200 are identical proteins with 23 repeating peptides. Twenty-one peptides contain potential N-glycosylation sites and these two glycoproteins differ only in their glycosylation patterns. Southern-blot analysis showed that a single-copy gene encodes both mRNAs. PGP200 is constitutively expressed, but the synthesis of SGP158 is totally dependent upon treatment of animals with the beta-agonist isoproterenol. The first 106-nucleotide sequence of cDNAs for PGP200 and SGP158, which corresponds to the 5'-untranslated region and sequence encoding the signal peptide, is highly conserved when compared with proline-rich protein and glutamine-rich protein gene sequences. Based on the nucleotide sequences of exon I, a phylogenetic tree was constructed for 35 members of these multigene families. The tree fits with the generally recognized phylogeny of mammalian orders. We propose that exon I sequences of the proline-rich protein and glutamine-rich protein multigene families are relatively new and are possibly generated through exon shuffling during evolution.
Asunto(s)
ADN Complementario/aislamiento & purificación , Familia de Multigenes , Péptidos/genética , Proteínas y Péptidos Salivales/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Exones , Glicosilación , Masculino , Datos de Secuencia Molecular , Péptidos/química , Dominios Proteicos Ricos en Prolina , Ratas , Ratas Sprague-Dawley , Proteínas Salivales Ricas en ProlinaRESUMEN
The liver is an attractive target tissue for gene therapy. Current approaches for hepatic gene delivery include retroviral and adenoviral vectors, liposome/DNA, and peptide/DNA complexes. This study describes a technique for direct injection of DNA into liver that led to significant gene expression. Gene expression was characterized in both rats and cats following injection of plasmid DNA encoding several different proteins. Luciferase activity was measured after injection of plasmid DNA encoding the luciferase gene (pCMVL), beta-galactosidase (beta-Gal) activity was evaluated in situ using plasmid DNA encoding Lac Z (pCMV beta), and serum concentration of secreted human alpha-1-antitrypsin was measured following injection of plasmid DNA encoding this protein (pRC/CMV-sHAT). Several variables, including injection technique, DNA dose, and DNA diluent, were investigated. Direct injection of pCMVL resulted in maximal luciferase expression at 24-48 hr. beta-Gal staining demonstrated that the majority of transfected hepatocytes were located near the injection site. Significant concentrations of human alpha-1-antitrypsin were detected in the serum of animals injected with pRC/CMV-sHAT. These findings demonstrate the general principle that direct injection of plasmid DNA into liver can lead to significant gene expression.
Asunto(s)
ADN/administración & dosificación , Expresión Génica , Hígado/metabolismo , Animales , Gatos , Terapia Genética , Humanos , Inyecciones , Luciferasas/biosíntesis , Plásmidos , Ratas , Ratas Sprague-Dawley , Células Tumorales Cultivadas , alfa 1-Antitripsina/biosíntesis , beta-Galactosidasa/biosíntesisRESUMEN
The critical physiological functions of the liver make hepatocytes important targets for therapeutic gene delivery. This study reports significant gene expression following direct injection of plasmid DNA into the livers of rats and cats. Transfection was characterized using luciferase and Lac Z expression from plasmids with the cytomegalovirus immediate early promoter/enhancer (CMV IE) or the Rous sarcoma virus long terminal repeat (RSV LTR). Dexamethasone treatment enhanced and prolonged transfected gene expression, possibly by activating gene expression. Southern analysis of total DNA extracted from liver at various times following injection detected persistent unintegrated plasmid DNA which maintained a prokaryotic methylation pattern. This study demonstrates the feasibility of direct DNA injection in the experimental analysis of hepatic gene expression in vivo.
Asunto(s)
ADN Recombinante , Dexametasona/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Hígado , Animales , Secuencia de Bases , Gatos , Células Cultivadas , Operón Lac , Hígado/citología , Hígado/efectos de los fármacos , Hígado/metabolismo , Luciferasas/genética , Masculino , Datos de Secuencia Molecular , Plásmidos , Ratas , Ratas Sprague-Dawley , Secuencias Repetitivas de Ácidos NucleicosAsunto(s)
Western Blotting/métodos , Membranas Artificiales , Polivinilos , Proteínas/análisis , Animales , Células Cultivadas , PorcinosRESUMEN
Six healthy subjects (3 males, 3 females) were studied to assess phasic inspiratory responses of upper airway (UA) and diaphragm muscles to electrocortical arousal independent of other potential respiratory stimulation. Transient electroencephalographic (EEG) arousal (abrupt EEG frequency shift > or = 3 s without awakening) was induced during supine stage 2 non-rapid-eye-movement (NREM) sleep with binaural tone bursts (0.5 s, 4 kHz, 25-95 dB). Electromyograms (EMG) of levator veli palatini (EMGlvp) and genioglossus (EMGgg) were obtained with intramuscular electrodes, and EMG of diaphragm (EMGdi) was obtained with esophageal electrodes. EMG signals were processed as moving time-averaged inspiratory activity over 100-ms windows. For each arousal, each of five consecutive postarousal breaths (R1-R5) was scored for peak inspiratory phasic EMG and normalized as percent averaged EMG of the three prearousal breaths for all muscles. After arousal, EMGlvp was increased for R1-R5 and EMGgg and EMGdi were increased for R1-R4. The increase in EMGlvp was greater than those of EMGgg and EMGdi for all response breaths. There was a significant increase in EMGlvp in all subjects, and EMGgg and EMGdi were significantly increased in three and two subjects, respectively. These data indicate that isolated transient electrocortical arousal is generally associated with phasic inspiratory recruitment of UA and diaphragm muscles in normal humans during NREM sleep; velopharyngeal muscle recruitment appears to be more consistent and of greater magnitude and duration than that of oropharyngeal muscle or diaphragm. We speculate that transient arousal from sleep may contribute to UA patency independent of chemical and mechanical respiratory stimuli.
Asunto(s)
Estimulación Acústica , Nivel de Alerta/fisiología , Corteza Cerebral/fisiología , Diafragma/fisiología , Músculos Faríngeos/fisiología , Sueño/fisiología , Adulto , Diafragma/inervación , Electroencefalografía , Electromiografía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Músculos Faríngeos/inervación , Polisomnografía , Reclutamiento Neurofisiológico/fisiología , Síndromes de la Apnea del Sueño/fisiopatologíaRESUMEN
Vitamin A (retinol) is required for the normal mucociliary differentiation of respiratory epithelium. A depletion of vitamin A promotes squamous cell metaplasia. To understand how vitamin A suppresses squamous cell differentiation, the expression of a squamous cell differentiation marker, the small proline-rich protein gene (spr1), was studied in cultured monkey tracheobronchial epithelial (TBE) cells. The expression of the spr1 gene was inhibited about 40 fold by retinol. The mRNA levels of the spr1 gene started to decline within 6 h of retinol treatment and reached a minimum level after 7 days. The inhibition by retinol was concentration dependent and did not require concurrent protein synthesis. The inhibition of the spr1 mRNA by retinol was not due to a decrease in the transcription rate of its gene but due to a decrease in its stability, as determined by nuclear run-on assays and mRNA half-life measurement, respectively. This result was further supported by a DNA transfection study using a chimeric construct containing the spr1 promoter region and the chloramphenicol acetyltransferase (CAT) reporter gene. The CAT activity in transfected cells was not inhibited by retinol. These results suggest that spr1 gene expression is posttranscriptionally down-regulated by retinol.
Asunto(s)
Bronquios/metabolismo , Regulación de la Expresión Génica , Proteínas/genética , Procesamiento Postranscripcional del ARN , Tráquea/metabolismo , Vitamina A/farmacología , Animales , Biomarcadores , Bronquios/citología , Diferenciación Celular/genética , Línea Celular , Proteínas Ricas en Prolina del Estrato Córneo , Regulación hacia Abajo , Células Epiteliales , Macaca mulatta , Proteínas de la Membrana , ARN Mensajero/metabolismo , Tráquea/citología , Transcripción GenéticaRESUMEN
In cell-free translations of RNA from primary cultures of pig trachea surface epithelial cells, we observed that a 20 kD proline-rich protein (sPRP) is induced during culturing (Biochem. Biophys. Res. Commun. 1990; 172:1304-1309). Subsequently, a cDNA encoding sPRP has been cloned from pig tracheal cell mRNA and sequenced. This cDNA shows a high similarity to cDNAs cloned from monkey tracheal cells cultured in vitamin A-free medium and from UV-irradiated human epidermal keratinocytes. Amino acid sequences from these cDNAs are exceptionally rich in proline, glutamine, cysteine, and lysine but contain no aromatic amino acids. Two repeats of 12 amino acids on the N-terminus are followed by multiple 8 amino acid repeats. When compared with monkey trachea and human keratinocyte cDNAs, the sPRP cDNA from pig trachea has an additional 24 bp nucleotide repeat. Antiserum raised to a synthetic peptide (23 amino acids) on the C-terminus of sPRP (C23-antiserum) reacted with the 20 kD sPRP in immunoprecipitations from cell-free translations. On Northern blot analysis, sPRP cDNA hybridized to RNAs of similar sizes in tracheal cells from cat, rabbit, and lamb. sPRP was not detected in tracheal cells that were cultured with 10(-9) M arotinoid. Since sPRP is considered a putative squamous cell differentiation marker, experiments using lung tumors were performed. sPRP mRNA levels were dramatically increased in squamous lung tumors that were induced by injecting hamsters with 4-(methylnitrosamino)- 1-(3-pyridyl)-1-butanone, a tobacco-specific nitrosamine. In situ hybridization with tissue sections prepared from these lung tumors revealed that cells around the keratin pearls contained high levels of sPRP mRNA.
Asunto(s)
Neoplasias Pulmonares/metabolismo , Biosíntesis de Péptidos , Péptidos , Prolina , Tráquea/metabolismo , Vitamina A/farmacología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Biomarcadores de Tumor/biosíntesis , Northern Blotting , Células Cultivadas , Cricetinae , ADN , Células Epiteliales , Haplorrinos , Humanos , Hibridación in Situ , Mesocricetus , Datos de Secuencia Molecular , Pruebas de Precipitina , Dominios Proteicos Ricos en Prolina , ARN Mensajero/análisis , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , PorcinosRESUMEN
The proline-rich proteins (PRPs) in mammalian salivary glands are encoded by tissue-specific multigene families whose members have diverged with respect to structure and regulation of expression. PRPs are expressed constitutively in humans, and comprise about [70%] of the total salivary proteins. Families of similar proteins are dramatically increased or induced in parotid and submandibular glands of rats, mice and hamsters by treatment with the [beta-] agonist isoproterenol. Feeding tannins to rats and mice mimics the effects of isoproterenol on the parotid glands. Salivary PRPs may constitute a defense mechanism against tannins and other polyhydroxylated phenols ingested. Putative transcriptional regulatory sequences have been identified in mouse PRP genes.
Asunto(s)
Péptidos/química , Péptidos/genética , Prolina/química , Prolina/genética , Proteínas y Péptidos Salivales/química , Proteínas y Péptidos Salivales/genética , Secuencia de Aminoácidos , Animales , Fenómenos Bioquímicos , Bioquímica , Regulación de la Expresión Génica , Humanos , Biología Molecular , Datos de Secuencia Molecular , Dominios Proteicos Ricos en ProlinaRESUMEN
An unusual expression of a putative squamous cell marker, small proline-rich protein (spr1), in mucociliary epithelial cells of conducting airways was demonstrated in a serum-free culture system. A cDNA clone was isolated from the cDNA library of monkey tracheobronchial epithelial (TBE) cells by differential hybridization. This cDNA clone, MT5, exhibited 98% homology to a DNA sequence obtained from human keratinocytes treated with either UV light or phorbol esters (T. Kartasova et al., 1988, Mol. Cell. Biol. 8:2195-2230). The predicted peptide of MT5 is unusual for its high content of proline (29%), glutamine (18%), and cysteine (9%) and its repeated PKVPEPC units. The level of spr1 mRNA in cultured cells was inhibited more than 90% by vitamin A. In contrast, phorbol 12-myristate 13-acetate (PMA) stimulated the level of spr1 mRNA by 3- to 8-fold. This differential regulation coincided with the effects of these chemicals on the cornification of cultured TBE cells. Using MT5 as a probe, we have localized the tracheal spr1 gene on the human chromosome 1 by a Southern blot analysis using a panel of human-rodent somatic cell hybrid DNAs. The gene was further sublocalized to bands q22-23 by in situ hybridization.
Asunto(s)
Bronquios/metabolismo , Cromosomas Humanos Par 1 , Regulación de la Expresión Génica , Proteínas/genética , Tráquea/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Biomarcadores , Northern Blotting , Southern Blotting , Bronquios/citología , Mapeo Cromosómico , Proteínas Ricas en Prolina del Estrato Córneo , ADN , Epitelio/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Cariotipificación , Macaca mulatta , Proteínas de la Membrana , Datos de Secuencia Molecular , Biosíntesis de Proteínas , Acetato de Tetradecanoilforbol/farmacología , Tráquea/citología , Vitamina A/farmacologíaRESUMEN
Alkaptonuria is an extremely rare, autosomal recessive disorder in which the metabolic enzyme homogentisic acid oxidase is deficient. A common sequelae is the subsequent accumulation of homogentisic acid in collagenous tissues, such as the sclera, nose and ear lobes. The blue-black pigmentation found in patients with alkaptonuria is called ochronosis. Another ocular sign includes amber-colored oil globulation within Bowmans membrane of the cornea.