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1.
Nurs Res Pract ; 2021: 8870394, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33898064

RESUMEN

Practical nursing skills are complex and involve technical, theoretical, and practical aspects, caring perspectives adjusted to both patient and circumstances, as well as ethical and moral considerations. Patients' length of stay in hospitals is decreasing, and more advanced patient treatment is conducted in primary healthcare settings. Hence, education and nursing skills need adjustment in line with the rapidly evolving field of practice. Studies emphasize a need to uncover whether the technical aspect of nursing skills, in general, is challenging in students' learning. The aim of this study was to explore students' perspectives on practical nursing skills and how they can best learn these. Three focus group interviews were conducted with registered nurse students and intellectual disability nurse students in their last semester (n = 11). Conventional, inductive content analysis in line with recommendations from Hsieh and Shannon was used to analyze the data. Two main categories with subcategories were identified: (1) the content of practical skills, with subcategories (a) human-to-human relations, (b) organizational competence, and (c) technical mastering and (2) building competence, with subcategories (a) need for supervision, (b) planning the learning situations, and (c) relevance for practice. Students experienced that practical skills did not only include technical aspects but also the ability to establish a relationship to the patient and to organize their working day. Supervising was assumed as essential both when training in the simulation center and in clinical placement, as well as planning of the training, respectively. Students experienced that some skills learned in the university college were less relevant in clinical practice and that certain skills were difficult to perform in practice due to the type of clinical placement. Hence, there is a need to review the approach to and content of practical nursing skills' learning in healthcare undergraduate programs, to prepare students for clinical practice, and to ensure that they build the competence needed in healthcare services.

2.
Structure ; 2(4): 293-308, 1994 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-8087556

RESUMEN

BACKGROUND: Lipases constitute a family of enzymes that hydrolyze triglycerides. They occur in many organisms and display a wide variety of substrate specificities. In recent years, much progress has been made towards explaining the mechanism of these enzymes and their ability to hydrolyze their substrates at an oil-water interface. RESULTS: We have determined the DNA and amino acid sequences for lipase B from the yeast Candida antarctica. The primary sequence has no significant homology to any other known lipase and deviates from the consensus sequence around the active site serine that is found in other lipases. We have determined the crystal structure of this enzyme using multiple isomorphous replacement methods for two crystal forms. Models for the orthorhombic and monoclinic crystal forms of the enzyme have been refined to 1.55 A and 2.1 A resolution, respectively. Lipase B is an alpha/beta type protein that has many features in common with previously determined lipase structures and other related enzymes. In the monoclinic crystal form, lipid-like molecules, most likely beta-octyl glucoside, can be seen close to the active site. The behaviour of these lipid molecules in the crystal structure has been studied at different pH values. CONCLUSION: The structure of Candida antarctica lipase B shows that the enzyme has a Ser-His-Asp catalytic triad in its active site. The structure appears to be in an 'open' conformation with a rather restricted entrance to the active site. We believe that this accounts for the substrate specificity and high degree of stereospecificity of this lipase.


Asunto(s)
Candida/enzimología , Proteínas Fúngicas/química , Lipasa/química , Conformación Proteica , Secuencia de Aminoácidos , Secuencia de Bases , Sitios de Unión , Clonación Molecular , Secuencia de Consenso , Cristalización , Cristalografía por Rayos X , Enlace de Hidrógeno , Modelos Moleculares , Datos de Secuencia Molecular , Solventes , Agua
3.
Oncogene ; 34(4): 424-35, 2015 Jan 22.
Artículo en Inglés | MEDLINE | ID: mdl-24469032

RESUMEN

S100A4 is implicated in metastasis and chronic inflammation, but its function remains uncertain. Here we establish an S100A4-dependent link between inflammation and metastatic tumor progression. We found that the acute-phase response proteins serum amyloid A (SAA) 1 and SAA3 are transcriptional targets of S100A4 via Toll-like receptor 4 (TLR4)/nuclear factor-κB signaling. SAA proteins stimulated the transcription of RANTES (regulated upon activation normal T-cell expressed and presumably secreted), G-CSF (granulocyte-colony-stimulating factor) and MMP2 (matrix metalloproteinase 2), MMP3, MMP9 and MMP13. We have also shown for the first time that SAA stimulate their own transcription as well as that of proinflammatory S100A8 and S100A9 proteins. Moreover, they strongly enhanced tumor cell adhesion to fibronectin, and stimulated migration and invasion of human and mouse tumor cells. Intravenously injected S100A4 protein induced expression of SAA proteins and cytokines in an organ-specific manner. In a breast cancer animal model, ectopic expression of SAA1 or SAA3 in tumor cells potently promoted widespread metastasis formation accompanied by a massive infiltration of immune cells. Furthermore, coordinate expression of S100A4 and SAA in tumor samples from colorectal carcinoma patients significantly correlated with reduced overall survival. These data show that SAA proteins are effectors for the metastasis-promoting functions of S100A4, and serve as a link between inflammation and tumor progression.


Asunto(s)
Inflamación/complicaciones , Metástasis de la Neoplasia , Proteínas S100/fisiología , Proteína Amiloide A Sérica/genética , Animales , Línea Celular Tumoral , Neoplasias del Colon/mortalidad , Receptores ErbB/fisiología , Humanos , Ratones , Especificidad de Órganos , Proteína de Unión al Calcio S100A4 , Proteína Amiloide A Sérica/fisiología
4.
Gene ; 73(1): 113-20, 1988 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-3072250

RESUMEN

Plasmids were constructed which contained two expression units encoding single-chain insulin precursors. Surprisingly, the total amount of insulin precursor produced was similar to that produced from plasmids containing a single expression unit. In this system, therefore, two expression cassettes can be brought to compete for the limited ability of the yeast cell for synthesis and secretion. Using genes encoding B(1-29)-A(1-21) and B(1-29)-Ala-Ala-Lys-A-(1-21), the slightly different precursors could be quantified individually after separation by high-performance liquid chromatography from the culture supernatant. The two-cassette system allowed a sensitive and well controlled comparison of parameters important for optimal expression of a heterologous gene in Saccharomyces cerevisiae. The system was used to compare two promoter constructions and also to evaluate the position of expression cassettes in the plasmid. Finally the codon usage in the gene to be expressed was found to influence its ability to compete for expression.


Asunto(s)
Genes , Insulina/genética , Plásmidos , Saccharomyces cerevisiae/genética , Secuencia de Aminoácidos , Secuencia de Bases , Southern Blotting , Cromatografía Líquida de Alta Presión , Codón , Humanos , Insulina/análisis , Insulina/biosíntesis , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Regiones Promotoras Genéticas , Proteínas Recombinantes/análisis , Regiones Terminadoras Genéticas
5.
FEBS Lett ; 212(2): 307-12, 1987 Feb 23.
Artículo en Inglés | MEDLINE | ID: mdl-3545904

RESUMEN

A yeast expression plasmid encoding a mini-proinsulin molecule was constructed and transformed into Saccharomyces cerevisiae. The plasmid encoded the sequence: B-Arg-Arg-Leu-Gln-Lys-Arg-A in which B represents the B-chain (30 amino acid residues) and A represents the A-chain (21 amino acid residues) of human insulin. The secreted peptides were shown to be a mixture of human insulin and des(B-30)human insulin. Thus, correct disulphide bridges can be established in proinsulin-like molecules devoid of a normal C-peptide region. Furthermore, the specificity of the yeast processing enzymes is so similar to the proinsulin converting enzymes in the human pancreatic beta-cell that it allows the processing of the mini-proinsulin to insulin.


Asunto(s)
Genes , Insulina/genética , Saccharomyces cerevisiae/genética , Transformación Genética , Secuencia de Aminoácidos , Humanos , Insulina/biosíntesis , Mapeo Peptídico , Plásmidos , Proteínas Recombinantes/aislamiento & purificación
6.
FEBS Lett ; 212(2): 302-6, 1987 Feb 23.
Artículo en Inglés | MEDLINE | ID: mdl-3028866

RESUMEN

Saccharomyces cerevisiae strains were transformed with plasmids coding for modified mating factor alpha 1 leader sequences followed by glucagon. Glucagon-containing peptides which were secreted into the fermentation broth were isolated and their amino acid sequences determined. The yeast strain transformed with the sequence coding for the complete mating factor alpha 1 leader sequence preceding the glucagon gene (MT556) secreted glucagon plus glucagon extended at its N-terminal by parts of the leader sequence. The yeast strain transformed with the sequence coding for a truncated mating factor alpha 1 leader sequence before the glucagon gene (MT615) secreted glucagon. These observations suggest that S. cerevisiae is a suitable vehicle for the efficient expression of plasmids coding for polypeptides similar to glucagon (e.g. VIP, secretin, GIP).


Asunto(s)
Genes Sintéticos , Genes , Glucagón/genética , Saccharomyces cerevisiae/genética , Transformación Genética , Secuencia de Aminoácidos , Secuencia de Bases , Enzimas de Restricción del ADN , Genes Fúngicos , Glucagón/biosíntesis , Factor de Apareamiento , Mutágenos , Péptidos , Plásmidos
7.
Mutat Res ; 106(2): 209-16, 1982 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-6185840

RESUMEN

The sensitivity to psoralen plus near-ultraviolet radiation (PUVA) was compared in a pair of E. coli strains differing at the acrA locus. Survival was determined for both bacteria and phage lambda. AcrA mutant cells were 40 times more sensitive than wild type to the lethal effect of PUVA. Free lambda phage exposed to PUVA survived as well when plated on acrA mutants as on wild type. In contrast, prophage lambda CI857 ind carried in lysogenic acrA strains was hypersensitive to PUVA. The enhanced sensitivity of bacterial and lambda DNA, when inside acrA cells, was paralleled by an increased photobinding of radiolabelled psoralens in the mutant. Binding was increased specifically to DNA rather than to nucleic acids in general. The difference in psoralen-binding ability determined by the acrA gene persisted after permeabilizing treatment of the cells. The results suggest that the acrA mutation causes an alteration specifically in the environment of the cellular DNA so as to allow increased intercalation and photobinding of psoralens.


Asunto(s)
Escherichia coli/efectos de los fármacos , Furocumarinas/farmacología , Bacteriófago lambda/efectos de los fármacos , Bacteriófago lambda/efectos de la radiación , ADN Bacteriano/metabolismo , ADN Viral/metabolismo , Escherichia coli/efectos de la radiación , Mutación , ARN Bacteriano/metabolismo , Rayos Ultravioleta
8.
Harv Bus Rev ; 79(3): 106-16, 165, 2001 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-11246918

RESUMEN

Most companies do a poor job of capitalizing on the wealth of expertise scattered across their organizations. That's because they tend to rely on centralized knowledge-management systems and technologies. But such systems are really only good at distributing explicit knowledge, the kind that can be captured and codified for general use. They're not very good at transferring implicit knowledge, the kind needed to generate new insights and creative ways of tackling business problems or opportunities. The authors suggest another approach, something they call T-shaped management, which requires executives to share knowledge freely across their organization (the horizontal part of the "T"), while remaining fiercely committed to their individual business unit's performance (the vertical part). A few companies are starting to use this approach, and one--BP Amoco--has been especially successful. From BP's experience, the authors have gleaned five ways that T-shaped managers help companies capitalize on their inherent knowledge. They increase efficiency by transferring best practices. They improve the quality of decision making companywide. They grow revenues through shared expertise. They develop new business opportunities through the cross-pollination of ideas. And they make bold strategic moves possible by delivering well-coordinated implementation. All that takes time, and BP's managers have had to learn how to balance that time against the attention they must pay to their own units. The authors suggest, however, that it's worth the effort to find such a balance to more fully realize the immense value of the knowledge lying idle within so many companies.


Asunto(s)
Personal Administrativo , Comercio/organización & administración , Desarrollo de Personal/métodos , Humanos , Conocimiento , Cultura Organizacional , Estados Unidos
9.
Harv Bus Rev ; 77(2): 106-16, 187, 1999.
Artículo en Inglés | MEDLINE | ID: mdl-10387767

RESUMEN

The rise of the computer and the increasing importance of intellectual assets have compelled executives to examine the knowledge underlying their businesses and how it is used. Because knowledge management as a conscious practice is so young, however, executives have lacked models to use as guides. To help fill that gap, the authors recently studied knowledge management practices at management consulting firms, health care providers, and computer manufacturers. They found two very different knowledge management strategies in place. In companies that sell relatively standardized products that fill common needs, knowledge is carefully codified and stored in databases, where it can be accessed and used--over and over again--by anyone in the organization. The authors call this the codification strategy. In companies that provide highly customized solutions to unique problems, knowledge is shared mainly through person-to-person contacts; the chief purpose of computers is to help people communicate. They call this the personalization strategy. A company's choice of knowledge management strategy is not arbitrary--it must be driven by the company's competitive strategy. Emphasizing the wrong approach or trying to pursue both can quickly undermine a business. The authors warn that knowledge management should not be isolated in a functional department like HR or IT. They emphasize that the benefits are greatest--to both the company and its customers--when a CEO and other general managers actively choose one of the approaches as a primary strategy.


Asunto(s)
Comercio/organización & administración , Difusión de Innovaciones , Gestión de la Información , Conocimiento , Consultores , Competencia Económica , Humanos , Servicios de Información/organización & administración , Relaciones Interprofesionales , Cultura Organizacional , Objetivos Organizacionales , Desarrollo de Personal , Estados Unidos
10.
Harv Bus Rev ; 78(5): 74-84, 199, 2000.
Artículo en Inglés | MEDLINE | ID: mdl-11143156

RESUMEN

Business incubators such as Hotbank, CMGI, and Idealab! are a booming industry. Offering office space, funding, and basic services to start-ups, these organizations have become the hottest way to nurture and grow fledgling businesses. But are incubators a fleeting phenomenon born of an overheated stock market, or are they an important and lasting way of creating value and wealth in the new economy? The authors argue that one type of incubator, called a networked incubator, represents a fundamentally new and enduring organizational model uniquely suited to growing businesses in the Internet economy. It shares certain features with other incubators--mainly, it fosters a spirit of entrepreneurship and offers economies of scale. But its key distinguishing feature is its ability to give start-ups preferential access to a network of potential partners. Such incubators institutionalize their networking--they have systems in place to encourage networking, helping start-ups, for example, to meet with potential business allies. That doesn't mean incubatees get preferential treatment; it means only that they have built-in access to partnerships that might not have existed without the incubator. Even with this advantage, however, networked incubators can easily follow the road to ruin. To avoid failure, they must create a portfolio of companies and advisers that their incubatees can leverage. That can be done by strategically investing in portfolio firms and by enlisting a large set of business allies. It can also be done by establishing connections and relationships that are anchored more to the incubator than to particular individuals.


Asunto(s)
Competencia Económica , Eficiencia Organizacional , Emprendimiento/organización & administración , Internet/organización & administración , Conducta Cooperativa , Creatividad , Humanos , Estados Unidos
11.
Ugeskr Laeger ; 155(47): 3823-7, 1993 Nov 22.
Artículo en Danés | MEDLINE | ID: mdl-8256382

RESUMEN

Renal failure is a frequent complication in multiple myeloma and it is present in about 50% of patients with newly-diagnosed multiple myeloma. Renal failure at the time of diagnosis has earlier been associated with a bad prognosis, but a better prognostic factor is the response to chemotherapy. In general, it is important to distinguish between 1) renal insufficiency at the time of diagnosis, 2) acute renal insufficiency and 3) chronic renal insufficiency developing during the course of the disease. The patients in the first two groups are treated with intensive therapy which is long-lasting (median four to six weeks). The improved function of the kidney is correlated to an improved survival. Patients developing chronic renal insufficiency late in the course of the disease should receive palliative therapy. The most important factors that provoke acute renal insufficiency are dehydration, hypercalcaemia and/or infection, but renal insufficiency is also provoked by the use of nephrotoxic drugs, hyperuricaemia and/or hyperviscosity. Chronic renal insufficiency is provoked by deposits of light chains, infiltration by plasma cells or deposits of amyloid. The treatment consists of elimination of the provoking factors and start of chemotherapy.


Asunto(s)
Lesión Renal Aguda/etiología , Fallo Renal Crónico/etiología , Mieloma Múltiple/complicaciones , Lesión Renal Aguda/diagnóstico , Lesión Renal Aguda/terapia , Humanos , Fallo Renal Crónico/diagnóstico , Fallo Renal Crónico/terapia , Mieloma Múltiple/fisiopatología , Pronóstico
12.
Ugeskr Laeger ; 155(19): 1436-41, 1993 May 10.
Artículo en Danés | MEDLINE | ID: mdl-8316969

RESUMEN

Aminoglycosides are among the most active antimicrobial agents against Gram-negative infections, but they also share the potential for oto- and nephrotoxicity. Animal studies have shown that dosing aminoglycosides once daily is more efficient and less nephrotoxic than the conventional multiple daily dosing regimens. Pharmacokinetic and microbiological data support this finding. Clinical trials confirm that once-daily dosing is more efficient and less toxic than multiple daily dosing. The two most important risk factors for nephrotoxicity seem to be the duration of aminoglycoside treatment and high trough levels of aminoglycoside. Netilmicin and amikacin are the drugs most often used in clinical trials of once-daily dosing regimens. Recommendations for once-daily dosing of netilmicin are given.


Asunto(s)
Antibacterianos/administración & dosificación , Aminoglicósidos , Animales , Antibacterianos/efectos adversos , Antibacterianos/economía , Antibacterianos/farmacocinética , Ensayos Clínicos como Asunto , Análisis Costo-Beneficio , Dinamarca , Oído Interno/efectos de los fármacos , Femenino , Humanos , Riñón/efectos de los fármacos , Masculino
14.
J Bacteriol ; 134(1): 71-5, 1978 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-649572

RESUMEN

The complexity of the genome of Micrococcus radiodurans was determined to be (2.0 +/- 0.3) X 10(9) daltons by DNA renaturation kinetics. The number of genome equivalents of DNA per cell was calculated from the complexity and the content of DNA. A lower limit of four genome equivalents per cell was approached with decreasing growth rate. Thus, no haploid stage appeared to be realized in this organism. The replication time was estimated from the kinetics and amount of residual DNA synthesis after inhibiting initiation of new rounds of replication. From this, the redundancy of terminal genetic markers was calculated to vary with growth rate from four to approximately eight copies per cell. All genetic material, including the least abundant, is thus multiply represented in each cell. The potential significance of the maintenance in each cell of multiple gene copies is discussed in relation to the extreme radiation resistance of M. radiodurans.


Asunto(s)
ADN Bacteriano/genética , Genes , Micrococcus/genética , ADN Bacteriano/análisis , ADN Bacteriano/biosíntesis , Cinética , Micrococcus/análisis , Micrococcus/crecimiento & desarrollo , Renaturación de Ácido Nucleico , Tolerancia a Radiación
15.
J Bacteriol ; 152(3): 976-82, 1982 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-6815165

RESUMEN

The processing of damaged DNA was altered in a mitomycin C-sensitive mutant (mtcA) of Micrococcus radiodurans. Even though the mutant retained resistance to 254-nm UV radiation, it did not, in contrast to the wild-type strain, show any excessive DNA degradation or cell death when incubated with chloramphenicol after sublethal doses of either UV light or mitomycin C. The results suggest the constitutive synthesis of an enzyme system responsible for wild-type proficiency in the repair of mitomycin C-induced damage. An alternative system able to repair damage caused by mitomycin C was demonstrated in the mtcA background. In this strain, additional damage inflicted upon the cellular DNA effected a massive rescue of cells previously inactivated by mitomycin C. Rescue was provoked by ionizing radiation, by UV light, or by simple alkylating agents. Cells treated with psoralen plus near-UV radiation could be rescued only when inactivation was due primarily to psoralen-DNA interstrand cross-links rather than to monoadducts. The rescue of inactivated cells was prevented in the presence of chloramphenicol. These results can be interpreted most readily in terms of an alternative repair system able to overcome DNA interstrand cross-links produced by mitomycin C or psoralen plus near-UV light, but induced only by the more abundant number of damages produced by radiation or simple alkylating agents.


Asunto(s)
Reparación del ADN , Furocumarinas/farmacología , Micrococcus/metabolismo , Mitomicinas/farmacología , Trioxsaleno/farmacología , Rayos Ultravioleta , Alquilantes/farmacología , Proteínas Bacterianas/biosíntesis , Cloranfenicol/farmacología , ADN Bacteriano/metabolismo , Micrococcus/genética , Micrococcus/efectos de la radiación , Mitomicina , Mutación
16.
J Bacteriol ; 141(1): 81-6, 1980 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-7354007

RESUMEN

Four proteins, alpha beta, gamma, and delta, preferentially synthesized in ultraviolet light-treated cells of Micrococcus radiodurans, were characterized in terms of their molecular weights and isoelectric points. Within the sublethal-dose range, the differential rate of synthesis for these proteins increased linearly with the inducing UV dose. The degree of induction reached 100-fold, and the most abundant protein beta, amounted to approximately 2% of the total newly synthesized protein after irradiation. Damage caused by ionizing radiation or by treatment with mitomycin C also provoked the synthesis of the four proteins. The proportions between the individual proteins, however, varied strikingly with the damaging agent. In contrast to treatments which introduced damage in the cellular deoxyribonucleic acid, the mere arrest of deoxyribonucleic acid replication, caused by nalidixic acid or by starvation for thymine, failed to elicit the synthesis of either protein. Repair of deoxyribonucleic acid damage requires that a number of versatile and efficient processes by employed. It is proposed that the induced proteins participate in deoxyribonucleic acid repair in M. radiodurans. Mechanisms are discussed which would allow a differentiated cellular response to damages of sufficiently distinctive nature.


Asunto(s)
Proteínas Bacterianas/biosíntesis , ADN Bacteriano/efectos de la radiación , Micrococcus/efectos de la radiación , Rayos Ultravioleta , Proteínas Bacterianas/análisis , Electrones , Micrococcus/metabolismo , Mitomicinas/farmacología , Ácido Nalidíxico/farmacología , Radiación Ionizante , Timina/metabolismo
17.
J Bacteriol ; 116(2): 588-92, 1973 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-4583242

RESUMEN

The rates of elongation of beta-galactosidase and its messenger ribonucleic acid (RNA) were estimated in a polyamine-deficient mutant of Escherichia coli through an analysis of the kinetics of enzyme induction. The chain growth of beta-galactosidase was calculated from the time required after the appearance of an amino terminal fragment of 60 amino acids (auto-alpha) until completed enzyme began to accumulate. The elongation rate of beta-galactosidase messenger RNA was estimated from the time after induction at which streptolydigen-resistant, enzyme-forming capacity first appeared. Upon polyamine starvation, the rate of polypeptide elongation slowed from 17 to 10 amino acids per s and the messenger RNA elongation rate decreased from 47 to 30 nucleotides per s. These reductions in polymerization rates were proportional to the decrease in cellular growth rate produced by polyamine starvation. It was concluded that, although it is quite unlikely that polyamine levels are involved in regulation of cell growth, they may be acting as cofactors in the synthesis of RNA or protein, or both.


Asunto(s)
Escherichia coli/metabolismo , Galactosidasas/metabolismo , Mutación , Extensión de la Cadena Peptídica de Translación/efectos de los fármacos , Poliaminas/farmacología , ARN Mensajero/metabolismo , Estabilidad de Medicamentos , Escherichia coli/enzimología , Cinética , Putrescina/farmacología , ARN Bacteriano/biosíntesis , ARN Mensajero/biosíntesis , Factores de Tiempo
18.
South Med J ; 90(1): 75-7, 1997 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-9003831

RESUMEN

Perineal hernia formation is an infrequent but well-recognized complication of major pelvic surgery. Various methods of perineal reconstruction have been reported. This report describes one technique of perineal hernia repair using a unilateral gracilis myocutaneous flap. The gracilis myocutaneous flap provides well-vascularized tissue that is useful in many situations requiring reconstruction of the pelvis and perineum, especially when the area has been irradiated.


Asunto(s)
Herniorrafia , Perineo/cirugía , Colgajos Quirúrgicos/métodos , Anciano , Carcinoma/complicaciones , Femenino , Hernia/etiología , Humanos , Neoplasias del Colon Sigmoide/complicaciones
19.
Yeast ; 6(2): 127-37, 1990.
Artículo en Inglés | MEDLINE | ID: mdl-2183521

RESUMEN

Mutants of Saccharomyces cerevisiae which lack the KEX2-encoded endopeptidase are unable to process proteolytically the mating factor alpha (MF alpha) propheromone produced from the chromosomal MF alpha 1 and MF alpha 2 genes (Julius et al., 1983). Overproduction of pheromone precursor from multiple, plasmid-borne MF alpha genes did, however, lead to the production of active MF alpha peptides in the absence of the KEX2 gene product. S. cerevisiae therefore must possess an alternative processing enzyme. The cleavage site of this enzyme appeared identical to that of the KEX2-encoded endopeptidase. To identify the gene responsible for the alternative processing, we have isolated clones which allowed production of mature MF alpha in a kex2-disrupted strain even from the chromosomal MF alpha genes. The gene isolated in this way was shown also to be essential for the KEX2-independent processing of propheromone overproduced from plasmid-borne MF alpha 1. The amino acid sequence deduced from the gene shows extensive homology to a number of aspartyl proteases including the PEP4 and BAR1 gene products from S. cerevisiae. In contrast to the BAR1 gene product, the novel aspartyl protease (YAP3 for Yeast Aspartyl Protease 3) contains a C-terminal serine/threonine-rich sequence and potential transmembrane domain similar to those found in the KEX2 gene product. The corresponding gene YAP3 was located to chromosome XII. The normal physiological role of the YAP3 gene product is not known. Strains disrupted in YAP3 are both viable and able to process the mating factor a precursor.


Asunto(s)
Ácido Aspártico Endopeptidasas , Endopeptidasas/genética , Péptidos/metabolismo , Precursores de Proteínas/metabolismo , Saccharomyces cerevisiae/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Endopeptidasas/metabolismo , Genes Fúngicos , Factor de Apareamiento , Datos de Secuencia Molecular , Procesamiento Proteico-Postraduccional , Saccharomyces cerevisiae/enzimología , Proteínas de Saccharomyces cerevisiae
20.
EMBO J ; 3(7): 1581-5, 1984 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-6204865

RESUMEN

One single glucoamylase gene could be identified in the chromosomal DNA of Aspergillus niger by Southern blot analysis. This glucoamylase gene was isolated from a genomic library of A. niger DNA. The glucoamylase gene is situated on a 2.5-kb EcoRI-EcoRV fragment and contains five intervening sequences in the coding region. One 169-bp intron is involved in differential mRNA processing leading to the two different glucoamylase enzymes G1 and G2; the other four introns are all very small ranging from 55 to 75 bp in length. One intron has a significant homology to the coding region which immediately follows, and it contains the internal conserved sequence TACTAAC, which is also found in yeast chromosomal gene introns, and is thought to participate in mRNA splicing. Two transcription initiation sites and a typical eukaryotic promoter region with TATAAT and CAAT boxes are located upstream from the gene.


Asunto(s)
Aspergillus niger/genética , Procesamiento Postranscripcional del ARN , Secuencia de Aminoácidos , Amilasas/genética , Aspergillus niger/enzimología , Secuencia de Bases , Regulación de la Expresión Génica , Genes , Operón
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