Immunization of mice with Lactobacillus casei expressing a beta-intimin fragment reduces intestinal colonization by Citrobacter rodentium.

Enteropathogenic Escherichia coli (EPEC) is a common cause of diarrhea in children from developing countries. Intimate adhesion of the bacteria to intestinal cells occurs via binding of the adhesin intimin to the TIR receptor exposed on cell surfaces. Here, Lactobacillus casei expressing a fragment of ß-intimin (L. casei-Int(cv)) was tested as mucosal vaccines in mice against intestinal colonization with the murine pathogen Citrobacter rodentium. Oral or sublingual immunization of C57BL/6 mice with L. casei-Int(cv) induced anti-Int(cv) IgA in feces but no IgG in sera. Conversely, anti-Int(cv) IgG was induced in the sera of mice after sublingual immunization with purified Int(cv). All vaccines were able to decrease C. rodentium recovery from feces. However, this reduction was more evident and sustained over time in mice immunized with L. casei-Int(cv) by the sublingual route. These mice also displayed an increase in interleukin 6 (IL-6) and gamma interferon (IFN-γ) secretion by spleen cells 10 days after infection. Additionally, oral or sublingual immunization of C3H/HePas mice, which are highly susceptible to C. rodentium infection, with L. casei-Int(cv) induced anti-Int(cv) antibodies and significantly increased survival after challenge. Immunohistological analysis of colon sections revealed that C. rodentium was located in deep fractions of the tissue from C3H/HePas mice immunized with L. casei whereas superficial staining was observed in colon sections from mice immunized with L. casei-Int(cv.) The results indicate that vaccines composed of L. casei expressing intimin may represent a promising approach and that the C3H/HePas infection model with C. rodentium can be used to evaluate potential vaccines against EPEC.

Análise da confiabilidade de um método de mensuração do ângulo de pico de torque ativo dos isquiossurais / Analysis of the reliability of a method for measuring hamstring active peak torque angle

INTRODUÇÃO: O comprimento muscular pode ser inferido através da determinação da relação comprimento-tensão do músculo. Essa relação é tradicionalmente investigada por meio da medida do torque máximo produzido pelo músculo e do ângulo em que esse torque é gerado. OBJETIVO: O presente estudo verificou a confiabilidade teste-reteste de um método de mensuração do ângulo de pico de torque ativo dos isquiossurais em jovens saudáveis. MÉTODO: Vinte e cinco indivíduos saudáveis (22,88 ± 1,67 anos) foram avaliados duas vezes em um intervalo de três semanas. Um dinamômetro isocinético foi utilizado no modo passivo para avaliar o torque passivo dos isquiossurais. A atividade muscular foi monitorada para garantir silêncio eletromio-gráfico. O dinamômetro foi utilizado no modo concêntrico para determinar o torque total dos isquiossurais. O torque ativo foi obtido subtraindo-se o torque passivo do torque total. O ângulo do pico de torque ativo foi utilizado para a análise. RESULTADO: Não houve diferença estatisticamente significativa entre as duas medidas realizadas (t= 1,009; p= 0,323). O Coeficiente de Correlação Intraclasse para os valores obtidos do ângulo de pico de torque ativo foi de 0,948 (p= 0,0001; IC 95 por cento 0,881 - 0,977). CONCLUSÃO: O presente estudo demonstrou que o método descrito é confiável para a quantificação deste ângulo, sugerindo que este método pode ser utilizado para avaliar mudanças da curva torque-ângulo promovidas por alterações de comprimento muscular.

INTRODUCTION: Muscle length can be inferred from the length-tension relationship of the muscle. This relationship is traditionally investigated by measuring the peak torque produced by the muscle and the angle at which it is generated. OBJECTIVE: The present study investigated the test-retest reliability of a method for measuring hamstring active peak torque angle in healthy young adults. METHOD: Twenty-five healthy individuals (22.88 ± 1.67 years) were assessed twice with an interval of three weeks. An isokinetic dynamometer was used in passive mode to assess hamstring passive torque. Muscle activity was monitored to ensure electromyographic silence. The dynamometer was also used in concentric mode to determine hamstring total torque. The active torque was obtained as the difference between total torque and passive torque. The active peak torque angle was used for the analysis. RESULTS: There was no significant difference between the two measurements (t= 1.009; p= 0.323). The intraclass correlation coefficient for the active peak torque angle values obtained was 0.948 (p= 0.0001; 95 percent CI: 0.883 - 0.977). CONCLUSION: This study has shown that the method described is reliable for the quantification of active peak torque angle, thus suggesting that this method can be used to evaluate shifts in the torque-angle curve produced by muscle length changes.

Members of the ethylene signalling pathway are regulated in sugarcane during the association with nitrogen-fixing endophytic bacteria.

Nitrogen-fixing bacteria have been isolated from sugarcane in an endophytic and beneficial interaction that promotes plant growth. In this work, for the first time, the involvement of ethylene signalling in this interaction was investigated by molecular characterizing members of this pathway in sugarcane. The expression pattern of a putative ethylene receptor (SCER1) and two putative ERF transcription factors (SCERF1 and SCERF2) show exclusive modulation in plants inoculated with the diazotrophic endophytes. The gene expression profile of SCER1, SCERF1, and SCERF2 is differentially regulated in sugarcane genotypes that can establish efficient or inefficient associations with diazotrophic micro-organisms, exhibiting high or low biological nitrogen fixation (BNF) rates, respectively. In addition, SCER1, SCERF1, and SCERF2 expression is different in response to interactions with pathogenic and beneficial micro-organisms. Taken together, that data suggest that SCER1, SCERF1, and SCERF2 might participate in specific ethylene signalling cascade(s) that can identify a beneficial endophytic association, modulating sugarcane responses toward the diazotrophic endophytes.

A mitogenic signal triggered at an early stage of vaccinia virus infection: implication of MEK/ERK and protein kinase A in virus multiplication.

Vaccinia virus (VV) triggers a mitogenic signal at an early stage of infection. VV-induced proto-oncogene c-fos mRNA with kinetics paralleling that stimulated by serum. The VV virokine, or vaccinia virus growth factor (VGF), was not crucial for c-fos induction because it was observed upon infection with the virokine-minus mutant VV (VGF(-)). Furthermore, c-fos expression did not require infectious virus particles, as it occurred even with UV-inactivated VV and was equally induced by the different multiplicities of infection, i.e. 1.0, 5.0, and 25.0. c-fos expression was preceded by VV-induced DNA binding activity and was mediated via the cis-acting elements serum response element (SRE), activating protein-1 (AP-1), and cAMP-response element (CRE). VV activated the protein kinases p42MAPK/ERK2 and p44MAPK/ERK1 and the transcription factor ATF1 in a time-dependent manner with kinetics that paralleled those of VV-stimulated DNA-protein complex formation. The mitogenic signal transmission pathways leading to c-fos activation upon VV infection were apparently mediated by the protein kinases MEK, ERK, and PKA. This assumption was based on the findings that: 1) c-fos transcript was down-regulated; 2) the SRE, AP-1, and CRE binding activities were significantly reduced; and 3) the activation of p42MAPK/ERK2, p44MAPK/ERK1, and ATF1 were drastically affected when the viral infections were carried out in the presence of specific protein kinase inhibitor. Moreover, the mutant VV (VGF(-)) was also able to activate ERK1/2. It is noteworthy that virus multiplication was equally affected by the same kinase inhibitors. Taken together, our data provide evidence that the early mitogenic signal triggered upon VV infection relies upon the activation of the protein kinases MEK, ERK, and PKA, which are needed for both signal transduction and virus multiplication.

Differential neuronal expression and projections of melanin-concentrating hormone (MCH) and MCH-gene-overprinted-polypeptide (MGOP) in the rat brain.

The rat melanin-concentrating hormone (MCH) gene may produce, through alternative splicing, either the precursor of MCH and neuropeptide EI, two neuropeptides coexpressed in the zona incerta (ZI) and lateral hypothalamus (LHA), or a putative protein we named previously MCH-gene-overprinted-polypeptide (MGOP). First, we investigated the distribution and relative expression of MCH and MGOP mRNA in the rat brain by Northern blotting, RT-PCR and in situ hybridization. MGOP gene transcripts were detected mainly in the hypothalamus only by RT-PCR. Second, different antisera were raised toward the C-terminus of MGOP and used to identify the translational products. In the rat brain, no MGOP-processed peptide could be detected based on RP-HPLC coupled to specific RIA. A polypeptide of 14 kDa was found in the secretory pathway of transfected monkey COS7 cells expressing recombinant MGOP. In the rat hypothalamus, a specific protein of 12 kDa was identified by Western blot analysis. Finally, distribution of MGOP-immunoreactivity (IR) was investigated in the rat brain. Colocalization studies demonstrated that 98% of the MGOP-expressing perikarya in ZI/LHA also synthesized MCH. In addition, numerous, strongly stained MGOP-containing neurons were encountered in the hypothalamic periventricular nucleus. Perikarya labelled with MGOP antiserum were also found scattered in the cortex, caudate putamen, amygdala and lateral septal nucleus. MCH was not detected in these MGOP-containing neurons. Strikingly, dense staining of terminals was observed with MGOP antiserum but not with MCH antibodies in the suprachiasmatic, ventromedial and arcuate nuclei, and also in the external layer of the median eminence. These results demonstrated that MGOP and MCH-IR overlapped in LHA/ZI but displayed a differential distribution in other areas. Based on this cerebral distribution, MGOP may act as a new secreted protein in regulating many neuroendocrine functions, such as nursing, feeding and growth control in associated behavioural components.

Protein domains involved in nuclear transport of Fos.

The protein encoded by the proto-oncogene c-fos is constitutively nuclear in most cell types analyzed. It has a predicted molecular weight of about 55 kDa. Proteins with a molecular weight above 40 kDa cannot enter the nucleus passively. Our interest was to study which regions in the protein are involved in the nuclear transport. We prepared a series of deletions and point mutations of the protein and cloned the mutated genes into a eukaryotic expression vector. Cos-1 cells were used to express the mutants transiently. Using indirect immunofluorescence we studied the subcellular localization, analyzing the percentage of cells containing the protein in the nucleus, the cytoplasm, or both locations. Our studies showed that the Fos protein contains several regions which can act independently to translocate the protein into the nucleus.

The genome of cowpox virus contains a gene related to those encoding the epidermal growth factor, transforming growth factor alpha and vaccinia growth factor.

Cowpox virus (CPV) is a member of the Orthopoxvirus genus and has the genetic capacity to encode a multitude of genes that interfere with the host inflammatory and immune response or modulate the physiological state of infected and non-infected cells. Among these CPV factors are receptors homologous to interferon and tumor necrosis factor receptors and also a viral cellular serine-proteinase analog. Here we describe the detection of a CPV gene that encodes a protein homologous to epidermal growth factor, transforming growth factor alpha and poxvirus growth factors, such as the vaccinia growth factor (VGF). The VGF and other poxvirus growth factors are produced early in the infection and are secreted into the medium where they bind to the EGF receptors, generating mytotic responses. The cowpox growth factor (CGF) gene was detected in three copies on the virus genome by PCR, and by northern and southern blot hybridization using VGF nucleotide sequences as primers and probes. The CPV gene has a strong nucleotide and predicted amino acid similarity with VGF, and is also produced early in the infection.

Biological activities of a human amniotic membrane interferon.

In order to characterize further the human amniotic membrane interferon (IFN-AM), an interferon antigenically unrelated to human IFN-alpha, -beta, and -gamma or TNF, we analysed its biological activities. Here, we present direct evidence of its ability to affect cell growth and to induce the IFN-stimulated genes (ISGs) 6-16 and 2'-5' oligoadenylate synthetase (OAS), in addition to its crossed anti-viral activity. The cellular growth arrest effect of IFN-AM was dose-dependent and paralleled that of IFN-beta. IFN-AM was also able to inhibit thymidine incorporation into DNA, similar to IFN-beta. The mRNA induction of 6-16 gene with IFN-AM treatment reached its highest level at 500 IU/ml and remained constant up to 2000 IU/ml. Conversely, 2'-5' OAS mRNA induction was dose-dependent, with the maximum level detected at 2000 IU/ml of IFN-AM treatment. The time course of mRNA accumulation by ISGs with IFN-AM (500 IU/ml) stimulation was also investigated. Gene induction reached a maximum at 16 h after IFN treatment for 2'-5' OAS and at 48 h for the 6-16 gene. IFN-AM and human IFN-alpha induced similar levels of the OAS enzyme. IFN-AM also showed small but significant activity in bovine cells. In conclusion, the amniotic membrane IFN here studied showed both anti-cellular activity and the ability to stimulate ISG-transcriptional activation in a similar manner to IFN-beta. In addition, IFN-AM was also as able to induce the expression of the enzyme 2'-5' OAS, as did IFN-alpha. Lastly, amniotic IFN showed a significant cross-species anti-viral activity, which was different from both human IFN-alpha and -beta. Taken together, these data strongly suggest that IFN-AM is a novel sub-type I IFN.

Morphological and molecular characterization of the poxvirus BeAn 58058.

BeAn 58058 virus (BAV) was isolated from an Oryzomis rodent in Brazil. BAV was shown to be antigenically related to another poxvirus also isolated in Brazil, the Cotia virus, but it remained ungrouped. Electron microscopy revealed that BAV has a typical poxvirus morphology. The Hind III DNA profile of BAV genome was similar with that of VV WR and Lister, but some differences in the profile were detected. We have also detected the presence of genes homologous to vaccinia virus (VV WR) genes in the genome of BAV. Genes related to vaccinia thymidine kinase (TK) gene and vaccinia growth factor (VGF) gene were found. The patterns of TK and VGF mRNA transcripts described for vaccinia virus infected cells were observed in BAV infected cells. Nucleotide sequence of BAV VGF homologous gene was similar to VV WR VGF sequences. This similarity was further seen when cross-hybridization of total genomes of BAV and VV was done. Polypeptide synthesis of BAV and vaccinia in infected cells also showed similar profiles. The genetic data was used to construct a phylogenetic tree where BAV and VV were placed at the same cluster. Based on our findings we propose that BAV is a vaccinia virus variant.

Presence of human immunodeficiency virus (HIV) and T-lymphotropic virus type I and II (HTLV-I/II) in a haemophiliac population in Belo Horizonte, Brazil, and correlation with additional serological results.

The aim of this study was to determine the prevalence of human immunodeficiency virus type 1 (HIV-1) and human T lymphotropic virus types I and II (HTLV-I/II) infections in 226 haemophiliac patients treated at Fundação Hemominas in Belo Horizonte, Minas Gerais State, Brazil, and to verify association with other serological results. Patients positive for HTLV-I/II had also a neurological, haematological and ophthalmological evaluation. Fundação Hemominas offers comprehensive care for all haemophiliac patients in Minas Gerais. Thirty-six (15.9%) of the 226 patients showed reactive results to HIV-1 [ELISA, Abbott, USA, confirmed by Western blot (WB), Cambridge Biotech, USA, and/or immunofluorescence, Fiocruz, Brazil] and 16 (7.1%) had reactive sera to HTLV-I/II (ELISA, Ortho). Eleven of these 16 (4.9%) were positive, 3/16 (1.3%) were indeterminate and 2/16 (0.9%) were negative in the HTLV WB (Cambridge Biotech). Neurological, haematological and ophthalmological examination of 9/16 patients revealed no abnormality suggestive of HTLV disease. Of the 16 patients reactive to HTLV-I/II ELISA test, six (37.5%) were also positive to HIV-1 (chi 2 = 5.92; P = 0.01). Seropositivity for HTLV-I/II and HIV-1 was associated with advancing age and positive results for hepatitis C virus (HCV), Chagas' disease (T. cruzi infection) and syphilis. No association between the presence of HTLV with type and severity of haemophilia and hepatitis B results was detected. The prevalence of antibodies against HIV-1 is approximately three times that of HTLV-I/II and a patient positive for HTLV-I/II had a significantly increased risk of being positive for HIV-1, HCV and T. cruzi.

The low proliferation rates of human amniotic cells are neither associated to deregulated proto-oncogenes' expression nor to the effect of IFN alpha 2.

Primary cultures of human amniotic membrane (PCHAM) cells display very low proliferation rates while their doubling times vary between 150 h and 210 h even after mitogenic stimuli. However, the pattern of proto-oncogenes (c-fos, c-myc and c-jun) expression in these cells, upon serum restimulation, resembled that of cell lines that display shorter population doubling times. Serum stimulation of quiescent PCHAM cells promoted a rapid and transient c-fos mRNA expression, which was detected within 10 min, reached maximal levels at 30 min and decreased to undetectable levels 2-3 h later. The levels of c-myc or c-jun mRNA increased within 10 min after serum restimulation, peaked at 3 h and decreased to intermediate levels thereafter. We also present evidence showing that IFN alpha 2 treatment of PCHAM cells had no effect on their population doubling times nor in c-fas, c-myc, or c-jun mRNA expression, under conditions in which induction of IFN-stimulated genes, such as 2'-5' oligo-adenylate synthetase (OAS) and 6-16 was observed. We conclude that the growth constraints observed with this cells are not directly associated with a negative cellular growth regulation exerted by IFN alpha 2, nor due to a deregulated proto-oncogenes' expression.

Distinct antigenic subtypes of human beta interferon can be distinguished by neutralization.

Different molecular configurations of human beta interferon were titrated with the standard reference antiserum of the National Institutes of Health (NIH) which had been prepared with natural beta fibroblast interferon in order to determine to what extent differences in these configurations would influence the neutralization of the antiviral action of interferon. Neutralization tests were carried out in Vero cells by diluting both interferon and antiserum. Encephalomyocarditis virus was employed as challenge virus. The neutralization titer was considered to have been reached when the effect of eight units of interferon was reduced to one. Two natural beta interferons prepared from fibroblasts and from amniotic membranes gave similar high titers. However, titers were reduced five-fold with recombinant interferons expressed in Escherichia coli, which do not contain carbohydrate, one with the natural sequence and a mutant with a single amino acid substitution (cysteine for serine). The NIH antiserum did not neutralize the effect of a protein fraction from amniotic membranes antigenically different from the human alpha, beta or gamma interferons but having the biological activity of interferon. We conclude that the carbohydrate moieties of human beta interferons are essential for their recognition by the NIH antiserum and that antibodies specific for human recombinant beta interferon, which does not contain carbohydrate, are needed.

Distinct antigenic subtypes of human beta-interferon can be distinguished by neutralization

Different molecular configurations of human beta interferon were titrated with the standard reference antiserum of the National Institutes of Health (NIH) which had been prepared with natural beta fibroblast interferon in order to determine to what extent differences in these configurations would influence the neutralization of the antiviral action of interferon. Neutralization tests were carried out in Vero cells by diluting both interferon and antiserum. Encephalomyocarditis virus was employed as challenge virus. The neutralization titer was considered to have been reached when the effect of eight units of interferon was reduced to one. Two natural beta interferons prepared from fibroblasts and from amniotic membranes gave similar high titers. However, titers were reduced five-fold with recombinant interferons expressed in Escherichia coli, which do not contain carbohydrate, one with the natural sequence and a mutant with a single amino acid substitution (cysteine for serine). The NIH antiserum did not neutralize the effect of a protein fraction from amniotic membranes antigenically different from the human alpha, beta, or gamma interferons but having the biological activity of interferon. We conclude that the carbohydrate moieties of human beta interferons are essential for their recognition by the NIH antiserum and that antibodies specific for human recombinant beta interferon, which does not contain carbohydrate, are needed.

Developmental expression of the arabidopsis cyclin gene cyc1At.

In eukaryotes, the control of cell cycle progression is exercised by heteromeric protein kinase complexes composed of a cell cycle-dependent, kinase-related subunit (Cdc2) and a cyclin subunit. To explore the possibility that cyclin transcription plays a role in the developmental regulation of cell division, we examined the spatial and temporal expression of a cyclin gene (cyc1At) in Arabidopsis. In root and shoot apical meristems and during embryogenesis, cyc1At expression is almost exclusively confined to dividing cells. A cell-specific pattern of cyc1At expression was noticed in root meristems. We examined the effects of induction of cell division of differentiated cells on cyc1At expression. During lateral root formation, induction of cyc1At expression is a very early event and was detected before anatomical modifications were visible. Treatment of roots with oryzalin, which blocks cell division in metaphase, did not inhibit the auxin induction of cyc1At, suggesting that induction of cyc1At expression precedes the completion of the first division cycle after induction of lateral roots. In tobacco protoplasts, an increase in cyc1At expression was observed only when cell division was induced. Together, the results suggest that Cyc1At accumulation in Arabidopsis is transcriptionally regulated and might be one of the limiting factors for the activation of cell division.

A protein phosphatase 1 from Arabidopsis thaliana restores temperature sensitivity of a Schizosaccharomyces pombe cdc25ts/wee1- double mutant.

There is increasing evidence that the mechanisms controlling the eukaryotic cell cycle are regulated by protein phosphorylation/dephosphorylation cascades. The catalytic subunit of the protein phosphatase 1 is implicated genetically and biochemically in the complex network that regulates mitosis. To investigate further the cell division in plants, we have isolated and characterized two full-length cDNAs from Arabidopsis thaliana, PP1A-At1 and PP1A-At2, encoding polypeptides highly homologous to known protein phosphatase 1 (PP1). DNA gel blot analysis suggests that the protein phosphatases 1 might form a small gene family in Arabidopsis. Northern analysis shows that transcripts are present in all plant organs. In cell cultures, the PP1 mRNA levels are differentially affected by treatment with drugs that block cell division. The expression of PP1A-At1 in a Schizosaccharomyces pombe cdc25ts/wee1- double-mutant strain restores temperature sensitivity, showing that the Arabidopsis phosphatase gene is capable of interacting with genes that regulate the fission yeast mitotic apparatus. However, the dis2-11 S. pombe strain, which has a cold-sensitive allele of the phosphatase 1 gene, is not rescued by expression of the PP1A-At1 gene, suggesting that the plant cDNA is not a functional homolog of the fission yeast gene.

[Pioneer experience with buflomedil in the form of retrograde venous pulse therapy for treatment of ischemia and severe extremity infections]. / Pioniererfahrungen mit Buflomedil in Form der retrograden venösen Pulstherapie für die Behandlung von Ischämie und schweren Extremitäteninfektionen.

Retrograde venous pulse therapy (RVP) allows faster healing of the infection, shortening of hospitalization time, and recovering of the chronic ischemia. RVP als has a dramatic effect on diabetic foot by passing the functional disorder of the microcirculation produced by diabetic neuropathy and micro-angiopathy, with important reduction of amputation rates in these patients. We credit these results to the very high concentration of Buflomedil provided by RVP in the target tissues, which allows the penetration and impregnation of other active substances (7).

Some biological properties of the human amniotic membrane interferon

Human amniotic interferon was investigated to define the species specificity of its antiviral action and compare its anti-cellular and NK cell stimulating activities with those of other human interferons. The antiviral effect was titrated in bovine (RV-IAL) and monkey (VERO) cells. Amniotic interferon exhibited, in bovine cells, 5% of the activity seen in monkey cells, while alpha interferon displayed 200%. No effect was detected with either beta or gamma interferon in bovine cells. Daudi cells were exposed to different concentrations of various interferons and the cell numbers were determined. The anticellular effect of the amniotic interferon reached its peak on the third day of incubation. Results suggested a higher activity for alpha and gamma interferons and a lower activity for beta when compared to amniotic interferon. Using total mononuclear cells as effector cells and K 562 as target cell in a 51Cr release assay, it was demonstrated that low concentrations of amniotic interferon consistently stimulated NK cell activity in cells derived from several donors, the results indicating a higher level of activity with this interferon than with alpha and beta interferons

The Arabidopsis functional homolog of the p34cdc2 protein kinase.

The p34cdc2 protein kinase is a key component of the eukaryotic cell cycle, which is required for G1 to S-phase transition and for entry into mitosis. Using a 380-base pair DNA fragment obtained by polymerase chain reaction amplification from an Arabidopsis thaliana flower cDNA library as a probe, we isolated and sequenced a cdc2-homologous cDNA from Arabidopsis. The encoded polypeptide has extensive homology with cdc2-like kinases. Furthermore, when expressed in a CDC28ts Saccharomyces strain, it partially restores the capacity to grow at 36 degrees C, indicating that the plant cDNA is a functional homolog of the p34cdc2 kinase. Genomic hybridization demonstrated that there is one copy of the cdc2 gene per Arabidopsis haploid genome. Using RNA gel blot analysis, we found that cdc2 mRNA is present in all plant organs.

Partial characterization of human amniotic membrane interferon

The molecular weight and the electrofocusing profile of human amniotic membrane interferon (IFN-AM) were determined. When submitted to gel filtration, IFN-AM showed a single 26-28 kDa component; in polyacrylamide gel electrophoresis one component of 19,500. In electrofocusing, IFN-AM displayed a terogeneity was reduced by previous treatment of IFN-AM with neuraminidase. IFN-AMis a siaglycoprotein similarto human beta IFN in terms of antigenicity but different from it in electrofocusing profile

The product of a fos-related gene, fra-1, binds cooperatively to the AP-1 site with Jun: transcription factor AP-1 is comprised of multiple protein complexes.

fra-1 encodes a serum-inducible protein (Fra-1) that is antigenically related to Fos. We have characterized Fra-1 expression in serum-stimulated cells using antibodies raised against several regions of this protein. Fra-1, expressed transiently in COS cells or in serum-stimulated rat fibroblasts, undergoes extensive post-translational modification, primarily by phosphorylation of serine residues. It is present in both the nucleus and the cytoplasm and participates in a protein complex with Jun. Using proteins synthesized in reticulocyte lysates, we have shown that Fra-1, like Fos, binds to the AP-1 recognition element cooperatively with Jun. A truncated Fra-1 protein that contains the leucine zipper region but not an adjacent basic amino acid domain, complexes with Jun in vitro but fails to bind AP-1 oligonucleotides. These results demonstrate that Fra-1 contributes to the DNA-binding activity ascribed to transcription factor AP-1.