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2.
Mech Dev ; 107(1-2): 13-23, 2001 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11520660

RESUMEN

Drosophila Spitz is a homolog of transforming growth factor alpha (TGF-alpha) and is an activating ligand for the EGF receptor (Egfr). It has been shown that Star is required for Spitz activity. Here we show that Star is quantitatively limiting for Spitz production during eye development. We also show that Star and Spitz proteins colocalize in Spitz sending cells and that this association is not coincident with the site of translation--consistent with a function for Star in Spitz processing or transmission. Finally, we have defined minimal sequences within both Spitz and Star that mediate a direct interaction and show that this binding can occur in vivo.


Asunto(s)
Proteínas de Drosophila , Drosophila/metabolismo , Proteínas de la Membrana/metabolismo , Células Fotorreceptoras de Invertebrados/metabolismo , Secuencia de Aminoácidos , Animales , Apoptosis , Drosophila/crecimiento & desarrollo , Factor de Crecimiento Epidérmico/química , Células HeLa , Humanos , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Datos de Secuencia Molecular , Mutación , Células Fotorreceptoras de Invertebrados/citología , Células Fotorreceptoras de Invertebrados/crecimiento & desarrollo , Unión Proteica , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/metabolismo , Recombinación Genética , Transducción de Señal , Transfección
4.
Mol Biol Cell ; 10(3): 649-64, 1999 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-10069809

RESUMEN

The fundamental process of nucleocytoplasmic transport takes place through the nuclear pore. Peripheral pore structures are presumably poised to interact with transport receptors and their cargo as these receptor complexes first encounter the pore. One such peripheral structure likely to play an important role in nuclear export is the basket structure located on the nuclear side of the pore. At present, Nup153 is the only nucleoporin known to localize to the surface of this basket, suggesting that Nup153 is potentially one of the first pore components an RNA or protein encounters during export. In this study, anti-Nup153 antibodies were used to probe the role of Nup153 in nuclear export in Xenopus oocytes. We found that Nup153 antibodies block three major classes of RNA export, that of snRNA, mRNA, and 5S rRNA. Nup153 antibodies also block the NES protein export pathway, specifically the export of the HIV Rev protein, as well as Rev-dependent RNA export. Not all export was blocked; Nup153 antibodies did not impede the export of tRNA or the recycling of importin beta to the cytoplasm. The specific antibodies used here also did not affect nuclear import, whether mediated by importin alpha/beta or by transportin. Overall, the results indicate that Nup153 is crucial to multiple classes of RNA and protein export, being involved at a vital juncture point in their export pathways. This juncture point appears to be one that is bypassed by tRNA during its export. We asked whether a physical interaction between RNA and Nup153 could be observed, using homoribopolymers as sequence-independent probes for interaction. Nup153, unlike four other nucleoporins including Nup98, associated strongly with poly(G) and significantly with poly(U). Thus, Nup153 is unique among the nucleoporins tested in its ability to interact with RNA and must do so either directly or indirectly through an adaptor protein. These results suggest a unique mechanistic role for Nup153 in the export of multiple cargos.


Asunto(s)
Núcleo Celular/metabolismo , Proteínas de Complejo Poro Nuclear , Proteínas Nucleares/metabolismo , Animales , Anticuerpos/metabolismo , Anticuerpos/farmacología , Transporte Biológico/efectos de los fármacos , Femenino , Productos del Gen rev/metabolismo , Carioferinas , Proteínas Nucleares/inmunología , Oocitos/efectos de los fármacos , Oocitos/metabolismo , Poli G/metabolismo , Poli U/metabolismo , ARN/metabolismo , Precursores del ARN/metabolismo , ARN Nuclear Pequeño/metabolismo , Xenopus
5.
Genes Dev ; 12(14): 2131-43, 1998 Jul 15.
Artículo en Inglés | MEDLINE | ID: mdl-9679058

RESUMEN

Activation of the Cyclin B/Cdc2 kinase complex triggers entry into mitosis in all eukaryotic cells. Cyclin B1 localization changes dramatically during the cell cycle, precipitously transiting from the cytoplasm to the nucleus at the beginning of mitosis. Presumably, this relocalization promotes the phosphorylation of nuclear targets critical for chromatin condensation and nuclear envelope breakdown. We show here that the previously characterized cytoplasmic retention sequence of Cyclin B1, responsible for its interphase cytoplasmic localization, is actually an autonomous nuclear export sequence, capable of directing nuclear export of a heterologous protein, and able to bind specifically to the recently identified export mediator, CRM1. We propose that the observed cytoplasmic localization of Cyclin B1 during interphase reflects the equilibrium between ongoing nuclear import and rapid CRM1-mediated export. In support of this hypothesis, we found that treatment of cells with leptomycin B, which disrupted Cyclin B1-CRM1 interactions, led to a marked nuclear accumulation of Cyclin B1. In mitosis, Cyclin B1 undergoes phosphorylation at several sites, a subset of which have been proposed to play a role in Cyclin B1 accumulation in the nucleus. Both CRM1 binding and the ability to direct nuclear export were affected by mutation of these phosphorylation sites; thus, we propose that Cyclin B1 phosphorylation at the G2/M transition prevents its interaction with CRM1, thereby reducing nuclear export and facilitating nuclear accumulation.


Asunto(s)
Proteínas Portadoras/metabolismo , Ciclina B/metabolismo , Péptidos y Proteínas de Señalización Intracelular , Carioferinas , Proteínas Nucleares/metabolismo , Receptores Citoplasmáticos y Nucleares , Animales , Sitios de Unión , Transporte Biológico , Ciclina B/genética , Ciclina B1 , Ácidos Grasos Insaturados/farmacología , Guanosina Difosfato/metabolismo , Células HeLa , Humanos , Ratones , Fosforilación , Ratas , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Xenopus , Xenopus laevis , Proteína de Unión al GTP ran , Proteína Exportina 1
7.
J Cell Biol ; 136(2): 241-50, 1997 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-9015297

RESUMEN

The 97-kD O-linked glycoprotein, Nup98, is a component of the Xenopus laevis nuclear pore complex and the only vertebrate GLFG nucleoporin identified (Powers, M.A., C. Macauley, F. Masiarz, and D.J. Forbes. 1995. J. Cell Biol. 128:721-736). We have investigated possible roles of xNup98 in the nucleocytoplasmic transport of proteins and RNAs by analyzing the consequences of injecting monospecific polyclonal antibodies to xNup98 into X. laevis oocytes. We show here that nuclear injection of anti-xNup98 inhibited the export of multiple classes of RNAs, including snRNAs, 5S RNA, large ribosomal RNAs, and mRNA. In contrast, the export of tRNA was unaffected. Injection of anti-xNup98 into the oocyte cytoplasm had no effect on export of any of the RNAs. Significantly, nuclear injection of anti-xNup98 antibodies did not inhibit import of either karyophilic proteins or snRNPs. This latter result is in agreement with our previous finding that Nup98 is not an essential element of the protein import pathway. Thus, Nup98 plays a role specifically in RNA export from the nucleus, and it appears to be an essential component of multiple RNA export pathways.


Asunto(s)
Núcleo Celular/metabolismo , Proteínas de la Membrana/fisiología , Proteínas de Complejo Poro Nuclear , Proteínas Nucleares/fisiología , ARN/metabolismo , Animales , Anticuerpos , Transporte Biológico , Proteínas de la Membrana/inmunología , Proteínas Nucleares/inmunología , Proteínas Nucleares/metabolismo , Oocitos , Pruebas de Precipitina , Caperuzas de ARN/metabolismo , ARN Mensajero/metabolismo , ARN Ribosómico 18S/metabolismo , ARN Ribosómico 28S/metabolismo , ARN Ribosómico 5S/metabolismo , ARN Nuclear Pequeño/metabolismo , ARN de Transferencia/metabolismo , Xenopus laevis
8.
Proc Natl Acad Sci U S A ; 94(26): 14394-9, 1997 Dec 23.
Artículo en Inglés | MEDLINE | ID: mdl-9405623

RESUMEN

Leucine-rich nuclear export signals (NESs) are recognized by the NES receptor exportin 1 and are central to the export of multiple shuttling proteins and RNAs. The export of messenger RNA in vertebrates was, however, thought to occur by a different pathway, because inhibition by injection of a synthetic Rev NES conjugate could not be demonstrated. Here we find that peptide conjugates composed of the NES of either protein kinase A inhibitor protein (PKI) or the HIV-1 Rev protein, when coupled to human serum albumin, are potent inhibitors of mRNA and small nuclear RNA export. These results provide direct evidence that mRNA export in vertebrates depends on interactions between an NES and its cognate NES receptors. PKI NES conjugates are significantly more efficient at inhibiting RNA export than are REV NES conjugates, indicating that different NESs may have different abilities to promote protein and RNA export. Surprisingly, an expected control conjugate containing the mutant Rev NES sequence M10 strongly inhibited the export of intronless dihydrofolate reductase mRNA. Nuclear injection of NES peptide conjugates led to mislocalization to the nucleus of 10-20% of the cytoplasmic Ran GTPase-binding protein (RanBP1) indicating that RanBP1 shuttles between the nucleus and the cytoplasm via an NES pathway. These results demonstrate that in vertebrates the export of mRNA, like that of small nuclear RNA, 5S rRNA, and transport factors such as RanBP1, employs NES-mediated molecular machinery.


Asunto(s)
Proteínas Portadoras/metabolismo , Productos del Gen rev/metabolismo , Carioferinas , ARN Mensajero/metabolismo , Receptores Citoplasmáticos y Nucleares , Secuencia de Aminoácidos , Animales , Transporte Biológico , Femenino , Productos del Gen rev/genética , Humanos , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Transfección , Xenopus laevis , Proteína Exportina 1
9.
Diabetes Care ; 19(11): 1294-301, 1996 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-8908400

RESUMEN

The scientific literature demonstrates that fat replacers have a reasonable certainty of no harm. Whether they help produce desired health outcomes, i.e., decreased risk of coronary heart disease and certain types of cancer related to excess fat intake, weight reduction, changes in lipid profile, improved glycemic control, etc., depends on how individuals use these foods to change food choices and eating behaviors. As Miller and Rolls conclude, ...the use of fat-replaced foods alone should not be expected to produce spontaneous improvements in weight management. Such improvements will still be dependent on long-term behavioral changes that include not only modifications in fat, but also modifications in overall energy intake and increase in energy expenditure. (53) Though it has not been studied, one may conjecture that encouraging people with diabetes to use foods with fat replacers to achieve nutrition management goals requires sufficient education, continuous counseling, and an individual's conscientious commitment and readiness to change food habits.


Asunto(s)
Diabetes Mellitus/dietoterapia , Dieta para Diabéticos , Dieta con Restricción de Grasas , Grasas de la Dieta , Sustitutos de Grasa , Ingestión de Energía , Sustitutos de Grasa/efectos adversos , Humanos , Fenómenos Fisiológicos de la Nutrición , Seguridad , Estados Unidos , United States Food and Drug Administration
10.
J Virol ; 70(9): 6296-303, 1996 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-8709257

RESUMEN

Peripheral blood mononuclear cells (PBMCs) infected with the oncogenic retrovirus bovine leukemia virus (BLV) produce virus when cultured briefly. BLV can be transmitted in cocultures to adherent susceptible cells, which become infected, express viral proteins, and fuse into multinucleated syncytia several days later. PBMCs from 3 of 10 BLV-infected sheep displayed a lifelong deficiency in induction of syncytium formation among indicator cells in culture, although large numbers of PBMCs synthesized viral transcripts or capsid protein. Since the infected, syncytium-deficient PBMCs were > or = 97% B cells, the deficiency could not be attributed to altered host cell tropism. The syncytium-deficient phenotype was recapitulated in newly infected sheep, demonstrating that this property is regulated by the viral genotype. The alteration in the BLV genome delayed but did not prohibit the establishment of BLV infection in vivo. Envelope glycoproteins were synthesized in syncytium-deficient PBMCs, translocated to the cell surface, and incorporated into virions. However, monoclonal antibodies specific for the BLV surface glycoprotein did not stain fixed PBMCs of the syncytium-deficient phenotype. Moreover, an animal with syncytium-deficient PBMCs had lower titers of neutralizing antibodies throughout the first 5 years of infection than an animal with similar numbers of infected PBMCs of the syncytium-inducing phenotype. The syncytium-deficient variant productively infected indicator cells at greatly reduced efficiency, showing that the alteration affects an early step in viral entry or replication. These results suggest that the alteration maps in the env gene or in a gene whose product affects the maturation or conformation, and consequently the function, of the envelope protein complex.


Asunto(s)
Cápside/biosíntesis , ADN Viral/sangre , Productos del Gen env/biosíntesis , Células Gigantes , Virus de la Leucemia Bovina/fisiología , Leucocitos Mononucleares/virología , Animales , Anticuerpos Antivirales , Linfocitos B/fisiología , Linfocitos B/virología , Bovinos , Células Cultivadas , Técnicas de Cocultivo , Genes env , Hibridación in Situ , Virus de la Leucemia Bovina/genética , Leucocitos Mononucleares/fisiología , Proteínas de la Membrana/biosíntesis , Proteínas de la Membrana/sangre , Pruebas de Neutralización , Fenotipo , Provirus/fisiología , ARN Viral/biosíntesis , Ovinos , Factores de Tiempo , Transcripción Genética
11.
J Cell Biol ; 128(5): 721-36, 1995 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-7876300

RESUMEN

Xenopus egg extracts provide a powerful system for in vitro reconstitution of nuclei and analysis of nuclear transport. Such cell-free extracts contain three major N-acetylglucosaminylated proteins: p200, p97, and p60. Both p200 and p60 have been found to be components of the nuclear pore. Here, the role of p97 has been investigated. Xenopus p97 was isolated and antisera were raised and affinity purified. Immunolocalization experiments indicate that p97 is present in a punctate pattern on the nuclear envelope and also in the nuclear interior. Peptide sequence analysis reveals that p97 contains a GLFG motif which defines a family of yeast nuclear pore proteins, as well as a peptide that is identical at 11/15 amino acids to a specific member of the GLFG family, NUP116. An additional peptide is highly homologous to a second sequence found in NUP116 and other members of the yeast GLFG family. A monoclonal antibody to the GLFG domain cross-reacts with a major Xenopus protein of 97 kD and polyclonal antiserum to p97 recognizes the yeast GLFG nucleoporin family. The p97 antiserum was used to immunodeplete Xenopus egg cytosol and p97-deficient nuclei were reconstituted. The p97-depleted nuclei remained largely competent for nuclear protein import. However, in contrast to control nuclei, nuclei deficient in p97 fail to grow in size over time and do not replicate their chromosomal DNA. ssDNA replication in such extracts remains unaffected. Addition of the N-acetylglucosaminylated nuclear proteins of Xenopus or rat reverses these replication and growth defects. The possible role(s) of p97 in these nuclear functions is discussed.


Asunto(s)
Proteínas de la Membrana/metabolismo , Membrana Nuclear/metabolismo , Proteínas de Complejo Poro Nuclear , Proteínas Nucleares/metabolismo , Proteínas de Saccharomyces cerevisiae , Secuencia de Aminoácidos , Animales , Transporte Biológico , Núcleo Celular/fisiología , Cromosomas/metabolismo , Replicación del ADN , Femenino , Técnica del Anticuerpo Fluorescente , Proteínas Fúngicas , Proteínas de la Membrana/química , Proteínas de la Membrana/inmunología , Datos de Secuencia Molecular , Membrana Nuclear/química , Membrana Nuclear/inmunología , Proteínas Nucleares/química , Proteínas Nucleares/inmunología , Proteínas de Unión al ARN , Secuencias Repetitivas de Ácidos Nucleicos , Análisis de Secuencia , Homología de Secuencia de Aminoácido , Xenopus
14.
J Am Diet Assoc ; 94(5): 498-500, 1994 May.
Artículo en Inglés | MEDLINE | ID: mdl-8176121
15.
Diabetes Educ ; 19(5): 419-30, 1993.
Artículo en Inglés | MEDLINE | ID: mdl-8137697

RESUMEN

This article reviews the three categories of fat replacers and characteristics of the individual products. Most importantly, this update provides strategies for staying abreast of current and future foods, and stimulates considerations for diabetes educators when advising clients about fat-modified foods. Creative techniques and tools also are presented for educating clients about these foods.


Asunto(s)
Diabetes Mellitus/dietoterapia , Grasas de la Dieta , Ciencias de la Nutrición/educación , Automonitorización de la Glucosa Sanguínea , Diabetes Mellitus/rehabilitación , Humanos , Planificación de Menú , Educación del Paciente como Asunto
16.
J Am Diet Assoc ; 93(7): 755-7, 1993 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-8320400

RESUMEN

If, as dietitians, we are to expand our role and influence on patient care and in clinical research (as described by the DCCT model), we need to examine our strengths, obtain expertise in our areas of interest, educate nondietitians and investigators about the benefits of using our skills in the clinical and research environments, and continue to develop our mentoring system. We need to identify our own personal challenges that will make us more effective leaders in health care. Also, we must acknowledge that the journey to reach these new goals will not always be easy, but is tremendously rewarding--personally and professionally.


Asunto(s)
Diabetes Mellitus/dietoterapia , Dieta para Diabéticos , Dietética , Investigación , Glucemia/análisis , Diabetes Mellitus/tratamiento farmacológico , Ingestión de Alimentos , Humanos , Insulina/uso terapéutico , Mentores , Modelos Teóricos
17.
Diabetes Educ ; 18(5): 393-400, 1992.
Artículo en Inglés | MEDLINE | ID: mdl-1296888

RESUMEN

This paper provides an historical documentation and discussion of events that have influenced diabetes nutritional management in recent years. Many factors have shaped the nutrition care that persons with diabetes receive today. Nutrition science research is part of the history, as are myriad discoveries, research, advanced technologies, and evolving health care systems. This review of the past four decades will contribute a perspective of how we have gotten to where we are today.


Asunto(s)
Diabetes Mellitus/historia , Dieta para Diabéticos/historia , Diabetes Mellitus/dietoterapia , Historia del Siglo XX , Humanos , Fenómenos Fisiológicos de la Nutrición
18.
J Virol ; 66(8): 4769-77, 1992 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-1378509

RESUMEN

Bovine leukemia virus (BLV) expression is mostly silent in peripheral blood mononuclear cells (PBMCs) of infected animals. However, when infected cells are cultured, they are stimulated to produce virus. We studied viral transcription in PBMCs taken from BLV-infected sheep because the pattern of transcriptional activation in these cells should closely mimic activation of virus expression within mononuclear cells in vivo. BLV transcription was activated as early as 30 min after PBMCs were cultured. Expression was characterized by early and late stages, each distinguished by a unique pattern of cytoplasmic RNAs. In early expression, cytoplasmic viral RNA was exclusively the doubly spliced tax/rex transcript, although all transcripts were present in the nucleus. Early expression gave way rapidly to late expression, in which all viral transcripts accumulated in the cytoplasm. The polyclonal B-cell activator lipopolysaccharide increased the amount of viral RNA by at least twofold but did not alter the pattern of transcription. The transition from early to late expression required new protein synthesis and was blocked by the inhibitor cycloheximide. This requirement reflects the essential role of the viral Rex protein in the transition, but synthesis of cellular factors may be required as well. These results provide the first demonstration of staged viral expression in lymphocytes naturally infected by either BLV or the closely related human T-cell leukemia virus (HTLV) and validate the model of BLV and HTLV gene expression that previously was derived from transfection experiments performed mainly in nonlymphoid cells.


Asunto(s)
Genes Reguladores , Genes Virales , Virus de la Leucemia Bovina/genética , Leucemia Experimental/microbiología , Linfocitos/microbiología , Transcripción Genética , Animales , Northern Blotting , Cápside/análisis , Cápside/genética , Núcleo Celular/fisiología , Células Cultivadas , Cinética , Virus de la Leucemia Bovina/crecimiento & desarrollo , Virus de la Leucemia Bovina/aislamiento & purificación , Poli A/genética , Poli A/aislamiento & purificación , ARN/genética , ARN/aislamiento & purificación , ARN Mensajero/genética , ARN Mensajero/aislamiento & purificación , ARN Mensajero/metabolismo , ARN Neoplásico/genética , ARN Neoplásico/aislamiento & purificación , ARN Viral/genética , ARN Viral/aislamiento & purificación , Ovinos , Activación Viral
19.
South Med J ; 84(10): 1217-20, 1991 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-1925723

RESUMEN

Rhodococcus equi is a gram-positive pleomorphic bacillus that has been identified as a life-threatening pulmonary pathogen in the immunocompromised host. Infection with R equi may go unrecognized by physicians unacquainted with its presentation and unaware of the organism's ability to mimic diphtheroids and to stain weakly positive with an acid-fast stain.


Asunto(s)
Infecciones por Actinomycetales/microbiología , Neumonía/microbiología , Rhodococcus equi/aislamiento & purificación , Infecciones por Actinomycetales/diagnóstico , Infecciones por Actinomycetales/inmunología , Adulto , Seropositividad para VIH , Humanos , Masculino , Infecciones Oportunistas/microbiología , Neumonía/diagnóstico
20.
J Virol ; 65(9): 4959-65, 1991 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-1651415

RESUMEN

Infection by bovine leukemia virus (BLV) is characterized by a long clinical latency after which some individuals develop B-cell tumors. The contributions of the viral regulatory proteins Tax and Rex during clinical latency and disease are incompletely understood. To learn about Rex expression in the host, we used a sensitive immunoprecipitation assay to detect Rex antibodies throughout the course of BLV infection in sheep. Sixty percent of the infected animals produced Rex antibodies in intermittent episodes. This pattern differed markedly from that of antibodies to virion structural proteins, which were maintained in all animals throughout infection. Only one of two animals that developed tumors had detectable Rex antibodies at the time, although the other had previously demonstrated an especially strong Rex antibody response. We examined the Rex response in the context of BLV infection by comparing it with the frequency of circulating mononuclear blood cells that could transcribe BLV RNA or produce infectious virus. Episodes of Rex antibody occurrence followed some but not all increases in the number of BLV-transcribing cells. Since the appearance of circulating antibodies requires that the intracellular Rex protein be available to serve as antigen, the episodic pattern of occurrence of Rex antibodies could result from intermittent killing by virus-specific cytotoxic cells. Fluctuations in titer that were observed during some episodes of Rex response could be due to antibody retention by antigen present in lymphoid tissue.


Asunto(s)
Anticuerpos Antivirales/inmunología , Productos del Gen rex/inmunología , Virus de la Leucemia Bovina/inmunología , Leucemia Experimental/inmunología , Animales , Clonación Molecular , Productos del Gen rex/genética , Periodicidad , Pruebas de Precipitina , Ovinos
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