RESUMO
Transcription by RNA polymerase II is accompanied by cyclic phosphorylation and dephosphorylation of the carboxy-terminal heptapeptide repeat domain (CTD) of its largest subunit. We have used deletion and point mutations in Fcp1p, a TFIIF-interacting CTD phosphatase, to show that the integrity of its BRCT domain, like that of its catalytic domain, is important for cell viability, mRNA synthesis, and CTD dephosphorylation in vivo. Although regions of Fcp1p carboxy terminal to its BRCT domain and at its amino terminus were not essential for viability, deletion of either of these regions affected the phosphorylation state of the CTD. Two portions of this carboxy-terminal region of Fcp1p bound directly to the first cyclin-like repeat in the core domain of the general transcription factor TFIIB, as well as to the RAP74 subunit of TFIIF. These regulatory interactions with Fcp1p involved closely related amino acid sequence motifs in TFIIB and RAP74. Mutating the Fcp1p-binding motif KEFGK in the RAP74 (Tfg1p) subunit of TFIIF to EEFGE led to both synthetic phenotypes in certain fcp1 tfg1 double mutants and a reduced ability of Fcp1p to activate transcription when it is artificially tethered to a promoter. These results suggest strongly that this KEFGK motif in RAP74 mediates its interaction with Fcp1p in vivo.
Assuntos
Fosfoproteínas Fosfatases/metabolismo , Saccharomyces cerevisiae/enzimologia , Fatores de Transcrição TFII , Fatores de Transcrição/química , Fatores de Transcrição/metabolismo , Motivos de Aminoácidos , Sequência de Aminoácidos , Sítios de Ligação , Regulação Fúngica da Expressão Gênica , Holoenzimas/química , Holoenzimas/genética , Holoenzimas/metabolismo , Espectroscopia de Ressonância Magnética , Metanossulfonato de Metila/farmacologia , Mutação , Fenótipo , Fosfoproteínas Fosfatases/genética , Fosforilação , Ligação Proteica , Estrutura Terciária de Proteína/genética , RNA Polimerase II/química , RNA Polimerase II/genética , RNA Polimerase II/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Sequências Repetitivas de Ácido Nucleico , Saccharomyces cerevisiae/citologia , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Fator de Transcrição TFIIB , Fatores de Transcrição/genética , Ativação TranscricionalRESUMO
The aim of the present study was to assess whether there are gender differences in (1) levels of extracellular serotonin (5-HT) in the forebrain, and (2) the effect on 5-HT of a reuptake inhibitor, paroxetine, or a releasing drug, fenfluramine. In vivo microdialysis was used to measure 5-HT in the hypothalamus of male and regularly cycling female rats. Hypothalamic 5-HT was significantly lower in estrous females (0.83 +/- 0.05 pg/sample, n=33) than in male rats (1.04 +/- 0.06 pg, n=38). Levels in diestrous females (0.98 +/- 0.09 pg, n=38) were not significantly different from males. Paroxetine (1 mg/kg) increased hypothalamic 5-HT in males, and diestrous and estrous females to approximately 2 pg/sample. However, the increase in hypothalamic 5-HT produced by a maximally effective dose of paroxetine (10 mg/kg) was significantly greater in male rats and during diestrous than during estrous. d,l-Fenfluramine (10 mg/kg) evoked an increase in extracellular 5-HT to approximately 15 pg/sample in all groups. A higher dose of d,l-fenfluramine (20 mg/kg) produced a significantly greater increase in hypothalamic 5-HT in males than in females during estrous or diestrous. These results are consistent with other evidence that during estrous, when rats are responding to peak levels of estrogen and progesterone, 5-HT release is decreased.
Assuntos
Estro/metabolismo , Hipotálamo/metabolismo , Serotonina/metabolismo , Animais , Diestro/efeitos dos fármacos , Diestro/metabolismo , Relação Dose-Resposta a Droga , Estro/efeitos dos fármacos , Espaço Extracelular , Feminino , Fenfluramina/farmacologia , Hipotálamo/efeitos dos fármacos , Masculino , Microdiálise/métodos , Paroxetina/farmacologia , Ratos , Ratos Sprague-Dawley , Caracteres SexuaisRESUMO
50 h) were detected for the transport and release of a model protein (ribonuclease A) compared with that for the translucent region which showed no lag time. The results highlight the importance of carefully controlling matrix formation to ensure reproducible transport and release characteristics from polymer matrices.
Assuntos
Sistemas de Liberação de Medicamentos , Ácido Hialurônico/administração & dosagem , Ribonuclease Pancreático/química , Difusão , Ribonuclease Pancreático/administração & dosagem , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The effects of polymer percent esterification and protein molecular weight on the diffusion of two model proteins, deoxyribonuclease (DNase) and ribonuclease A (RNase A), through and from partially esterified hyaluronic acid membranes were compared. The permeability of the polymer membranes was inversely related to the degree of polymer esterification and the molecular weight of the protein. Transport rates of proteins through the membranes decreased dramatically over narrow ranges of polymer esterification. As expected, the apparent diffusivity of the larger protein in the polymer matrix was more sensitive to changes in membrane hydration than that of the smaller protein. These observations demonstrated the dependence of the mobility of large molecular weight proteins on polymer hydration and chain relaxation. The relationship between protein diffusion through and release from the modified hyaluronate matrices was also investigated using RNase A as a model. The release profiles from fully esterified membranes showed lag behavior and varied with protein load and hyaluronate hydrolysis rates, while release from less esterified membranes was rapid and independent of polymer esterification or hydrolysis. Potential applications of modified hyaluronate matrices in the controlled delivery of proteins are discussed.
Assuntos
Desoxirribonucleases/química , Ácido Hialurônico/química , Membranas Artificiais , Ribonuclease Pancreático/química , Preparações de Ação Retardada , Difusão , Estabilidade Enzimática , Ésteres/química , Cinética , Peso Molecular , Permeabilidade , Temperatura , Timidina/químicaRESUMO
The team functioning assessment tool (TFAT) has been shown to be a reliable behavioral marker tool for assessing nontechnical skills that are critical to the success of ward-based healthcare teams. This paper aims to refine and shorten the length of the TFAT to improve usability, and establish its reliability and construct validity. Psychometric testing based on 110 multidisciplinary healthcare teams demonstrated that the TFAT is a reliable and valid tool for measuring team members' nontechnical skills in regards to Clinical Planning, Executive Tasks, and Team Functioning. Providing support for concurrent validity, high TFAT ratings were predicted by low levels of organizational constraints and high levels of group potency. There was also partial support for the negative relationships between time pressure, leadership ambiguity, and TFAT ratings. The paper provides a discussion on the applicability of the tool for assessing multidisciplinary healthcare team functioning in the context of improving team effectiveness and patient safety for ward-based hospital teams.
Assuntos
Atitude do Pessoal de Saúde , Relações Interprofissionais , Erros Médicos/prevenção & controle , Equipe de Assistência ao Paciente/normas , Segurança do Paciente/normas , Psicometria/instrumentação , Adulto , Comportamento Cooperativo , Análise Fatorial , Humanos , Pessoa de Meia-Idade , Estudos Multicêntricos como Assunto , Observação , Variações Dependentes do Observador , Equipe de Assistência ao Paciente/organização & administração , Recursos Humanos em Hospital , Queensland , Reprodutibilidade dos TestesRESUMO
Bacteriophage T4 capsids seem to be assembled on E. coli protoplasmic membranes. This process seems to involve "lumps" of head protein, which convert to tau particles, which in turn give rise to empty heads. The empty heads leave the bacterial membrane and are then filled with DNA in the central region of the cell. T4 gene 16 and 17 products appear to be necessary for head filling.
Assuntos
Colífagos/crescimento & desenvolvimento , Escherichia coli , Membrana Celular/metabolismo , Colífagos/metabolismo , DNA Viral/metabolismo , Escherichia coli/metabolismo , Microscopia Eletrônica , Morfogênese , MutaçãoRESUMO
The genes that encode the subunits of the Clp protease of Escherichia coli, clpA and clpP, appear to be regulated differently from each other. The clpA gene does not seem to be under heat shock control (Y. S. Katayama, S. Gottesman, J. Pumphrey, S. Rudikoff, W. P. Clark, and M. R. Maurizi, J. Biol. Chem. 263:15226-15236, 1988). In contrast, the level of ClpP protein was increased in rpoH+ cells but not in null rpoH cells after an upshift in temperature from 17 to 43 degrees C. The level of ClpP protein in a null dnaK strain was also elevated relative to the level of ClpP protein in an otherwise isogenic dnaK+ strain. In two-dimensional gels, the ClpP protein was located in the position of the previously unidentified heat shock protein F21.5. No protein spot corresponding to F21.5 was present in two-dimensional gels of a null clpP strain. The clpP gene, therefore, appears to be a heat shock gene, expressed in a sigma 32-dependent manner and negatively regulated by DnaK; the product of clpP is the previously unidentified heat shock protein F21.5.
Assuntos
Escherichia coli/genética , Genes Bacterianos , Proteínas de Choque Térmico/genética , Serina Endopeptidases/genética , Fator sigma/metabolismo , Proteases Dependentes de ATP , Eletroforese em Gel Bidimensional , Escherichia coli/enzimologia , Genótipo , Proteínas de Choque Térmico/biossíntese , Proteínas de Choque Térmico/isolamento & purificação , Substâncias Macromoleculares , Regiões Promotoras Genéticas , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/isolamento & purificação , Serina Endopeptidases/biossíntese , Serina Endopeptidases/isolamento & purificaçãoRESUMO
Escherichia coli bacteria produce at least one 70 kD stress protein, the product of the dnaK gene. We have compared the rates of degradation of different types of abnormal proteins in null Ion E. coli with a partial deletion of the dnaK gene with the rates observed in null Ion dnaK+ cells. We have found that both canavanyl proteins and puromycyl polypeptides are degraded more slowly in the null dnaK mutants than in the dnaK+ strain. However, a temperature-sensitive mutant LacI protein is degraded more rapidly in the null dnaK strain. The stability of this temperature-sensitive LacI protein was also examined in detail under various other conditions.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Escherichia coli , Escherichia coli/genética , Genes Bacterianos , Genes , Proteínas de Choque Térmico HSP70 , Proteínas de Choque Térmico/genética , Mutação , Proteínas/metabolismo , Escherichia coli/metabolismo , Genótipo , Cinética , beta-Galactosidase/genéticaRESUMO
Escherichia coli mutants were isolated that supported the growth of a lambda Ots and, in at least one case, a lambda Bts phage at the normally nonpermissive temperature of 39 degrees C. In one such strain, Ots and Bts suppression ability appeared to be a function of the guaB gene. Ots suppression by the mutant guaB strain was prevented if high levels of guanine or xanthine were present in the medium. No other base had any effect on Ots suppression in this strain. Other strains carrying spontaneous mutations resulting in guanine or xanthine auxotrophy (guaA or guaB lesions, respectively) all allowed lambda Ots replication at 39 degrees C; Ots suppression in these strains was also abolished by addition of guanine to the medium. Thus, reduced intracellular guanine levels resulting from guaA or guaB mutations appeared to suppress the inability of lambda Ots and, at least in some cases, Bts bacteriophage to form plaques at 39 degrees C. In burst size experiments, a guaB mutant produced a larger phage yield per infected cell of both lambda Ots and lambda O+ phage at 39 degrees C than did a similar guaB+ strain. It appeared that a lower-than-normal level of guanine (or a guanine derivative) in these cells may permit unusually efficient lambda replication. The fact that O+ and lambda Ots bursts in the guaB mutant were reduced significantly by addition of exogenous guanine to the medium is consistent with this suggestion. Another strain that suppresses the Ots allele has no known auxotrophic requirements, and suppression in this strain was unaffected by addition of guanine to the medium; however, addition of cytidine to the medium specifically eliminated Ots suppression in this strain. The mutation responsible for allowing Ots replication in this strain is unknown.
Assuntos
Bacteriófago lambda/fisiologia , Escherichia coli/genética , Genes Bacterianos , Supressão Genética , Citidina/farmacologia , Escherichia coli/fisiologia , Genes Virais , Guanina/farmacologia , Mutação , Fenótipo , Recombinases Rec A/genética , Temperatura , Ensaio de Placa Viral , Proteínas Virais/genética , Replicação Viral , Xantina , Xantinas/farmacologiaRESUMO
Extracts made from Escherichia coli null dnaK strains contained elevated levels of ATP-dependent proteolytic activity compared with levels in extracts made from dnaK+ strains. This ATP-dependent proteolytic activity was not due to Lon, Clp, or Alp-associated protease. Comparison of the levels of ATP-dependent proteolytic activity present in lon rpoH dnaK mutants and in lon rpoH dnaK+ mutants showed that the level of ATP-dependent proteolytic activity was elevated in the lon rpoH dnaK mutant strain. These findings suggest that DnaK negatively regulates a new ATP-dependent proteolytic activity, independently of sigma 32. Other results indicate that an ATP-dependent proteolytic activity was increased in a lon alp strain after heat shock. It is not yet known whether the same protease is associated with the increased ATP-dependent proteolytic activity in the dnaK mutants and in the heat-shocked lon alph strain.
Assuntos
Trifosfato de Adenosina/fisiologia , DNA Circular/fisiologia , Proteínas de Escherichia coli , Escherichia coli/fisiologia , Protease La , Serina Endopeptidases/deficiência , Proteases Dependentes de ATP , DNA de Cinetoplasto , Regulação Bacteriana da Expressão Gênica , Proteínas de Choque Térmico/fisiologia , HidróliseRESUMO
The product of gene 31 is normally required for assembly of the T4 capsid. Two mutations that each bypass that requirement are shown to be located at separate sites in gene 23, which encodes the major structural protein of the capsid. A second phenotypic effect that characterizes both bypass31 mutant strains is the ability to multiply in host-defective strains, such as hdB3-1 and groEL mutants, on which wild-type T4 is unable to assemble capsids. The genetic data indicate that both phenotypic effects are due to the bypass31 mutation. Elimination of the requirement for both the phage protein, gp31, and the host protein, GroEL, by either of two single mutations in gene 23 indicates that GroEL and gp31 are normally needed to interact with gp23 in capsid assembly of wild-type T4.
Assuntos
Replicação do DNA , Escherichia coli/genética , Genes Virais , Mutação , Fagos T/genética , Raios Ultravioleta , Replicação do DNA/efeitos da radiação , Escherichia coli/efeitos da radiação , Frequência do Gene , Fenótipo , Fagos T/efeitos da radiação , Replicação Viral/efeitos da radiaçãoRESUMO
T4 bacteriophage mutants called bypass31 (byp31) that specifically suppress gene 31 amber mutations have been isolated and characterized. The mechanism by which the byp31 mutation, byp31-1, suppresses gene 31 nonsense mutations does not involve synthesis of gp31 or of a particular gp31 fragment; furthermore, the byp31 allele suppresses all nonsense mutations in gene 31 that have been tested. We detect no unusual properties among the T4 particles made in su- cells by the T4amN54byp31-1 double mutant. These virions, made in the absence of gp31, show normal heat sensitivity, normal sensitivity to osmotic shock, and normal morphology. Specific different gene 31 missense mutants are able to form plaques with high efficiencies on the following two types of host defective cells: (i) Escherichia coli groEL (Tilly et al., Proc. Natl. Acad. Sci. U.S.A. 78:1629-1633, 1981) mutants that block T4 capsid assembly and (ii) E. coli rho mutants in which T4+ heads are assembled, but in which tail production and DNA synthesis are blocked. (Note that not all rho mutants block T4 production [G. Binkowski and L. D. Simon, p. 342-350, in C. K. Mathews, E. M. Kutter, G. Mosig, and P. B. Berget, ed., Bacteriophage T4, 1983]; T4 is able to replicate in rho mutants such as rho ts15, whose principal defect is that they fail to terminate transcription.) The byp31-1 allele permits production of T4 particles in E. coli groEL host-defective mutants, but not in E. coli rho host mutants.
Assuntos
DNA Viral/isolamento & purificação , Escherichia coli/genética , Genes Virais , Genes , Mutação , Fagos T/genética , Proteínas Virais/genética , Replicação do DNA , DNA Viral/genética , Especificidade da Espécie , Supressão Genética , Proteínas Virais/análise , Replicação ViralRESUMO
A bacteriophage T4 gene which functions to inhibit Escherichia coli Lon protease has been identified. This pin (proteolysis inhibition) gene was selected for its ability to support plaque formation by a lambda Ots vector at 40 degrees C. Southern blot experiments indicated that this T4 gene is included within the 4.9-kilobase XbaI fragment which contains gene 49. Subcloning experiments showed that T4 gene 49.1 (designated pinA) is responsible for the ability of the Ots vector to form plaques at 40 degrees C. Deficiencies in Lon protease activity are the only changes known in E. coli that permit lambda Ots phage to form plaques efficiently at 40 degrees C. lon+ lysogens of the lambda Ots vector containing pinA permitted a lambda Ots phage to form plaques efficiently at 40 degrees C. Furthermore, these lysogens, upon comparison with similar lysogens lacking any T4 DNA, showed reduced levels of degradation of puromycyl polypeptides and of canavanyl proteins. The lon+ lysogens that contained pinA exhibited other phenotypic characteristics common to lon strains, such as filamentation and production of mucoid colonies. Levels of degradation of canavanyl proteins were essentially the same, however, in null lon lysogens which either contained or lacked pinA. We infer from these data that the T4 pinA gene functions to block Lon protease activity; pinA does not, however, appear to block the activity of proteases other than Lon that are involved in the degradation of abnormal proteins.
Assuntos
Desoxirribonucleases de Sítio Específico do Tipo II , Proteínas de Escherichia coli , Escherichia coli/enzimologia , Proteínas de Choque Térmico , Inibidores de Proteases/farmacologia , Protease La , Serina Endopeptidases/metabolismo , Fagos T/genética , Proteínas Virais/genética , Proteases Dependentes de ATP , Bacteriófago lambda/crescimento & desenvolvimento , Sequência de Bases , Clonagem Molecular , Meios de Cultura , Enzimas de Restrição do DNA , DNA Viral/genética , Eletroforese em Gel de Ágar , Genes Virais , Vetores Genéticos , Lisogenia , Dados de Sequência Molecular , Puromicina/análogos & derivados , Puromicina/metabolismo , Ensaio de Placa Viral , Proteínas Virais/metabolismoRESUMO
Transcription of the clpP-clpX operon of Escherichia coli leads to the production of two different sizes of transcripts. In log phase, the level of the longer transcript is higher than the level of the shorter transcript. Soon after the onset of carbon starvation, the level of the shorter transcript increases significantly, and the level of the longer transcript decreases. The longer transcript consists of the entire clpP-clpX operon, whereas the shorter transcript contains the entire clpP gene but none of the clpX coding sequence. The RpoH protein is required for the increase in the level of the shorter transcript during carbon starvation. Primer extension experiments suggest that there is increased usage of the sigma(32)-dependent promoter of the clpP-clpX operon within 15 min after the start of carbon starvation. Expression of the clpP-clpX operon from the promoters upstream of the clpP gene decreases to a very low level by 20 min after the onset of carbon starvation. Various pieces of evidence suggest, though they do not conclusively prove, that production of the shorter transcript may involve premature termination of the longer transcript. The half-life of the shorter transcript is much less than that of the longer transcript during carbon starvation. E. coli rpoB mutations that affect transcription termination efficiency alter the ratio of the shorter clpP-clpX transcript to the longer transcript. The E. coli rpoB3595 mutant, with an RNA polymerase that terminates transcription with lower efficiency than the wild type, accumulates a lower percentage of the shorter transcript during carbon starvation than does the isogenic wild-type strain. In contrast, the rpoB8 mutant, with an RNA polymerase that terminates transcription with higher efficiency than the wild type, produces a higher percentage of the shorter clpP-clpX transcript when E. coli is in log phase. These and other data are consistent with the hypothesis that the shorter transcript results from premature transcription termination during production of the longer transcript.
Assuntos
Adenosina Trifosfatases/genética , Escherichia coli/enzimologia , Regulação Bacteriana da Expressão Gênica , Regulação Enzimológica da Expressão Gênica , Óperon , RNA Bacteriano/biossíntese , Serina Endopeptidases/genética , Fator sigma , ATPases Associadas a Diversas Atividades Celulares , Carbono/metabolismo , Meios de Cultura , RNA Polimerases Dirigidas por DNA/genética , RNA Polimerases Dirigidas por DNA/metabolismo , Endopeptidase Clp , Escherichia coli/genética , Proteínas de Escherichia coli , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Chaperonas Moleculares , Regiões Promotoras Genéticas , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transcrição Gênica , Ativação TranscricionalRESUMO
Mating type switching in Saccharomyces cerevisiae initiates when Ho endonuclease makes a double-stranded DNA break at the yeast MAT locus. In this report, we characterize the fundamental biochemical properties of Ho. Using an assay that monitors cleavage of a MAT plasmid, we define an optimal in vitro reaction, showing in particular that the enzyme has a stringent requirement for zinc ions. This suggests that zinc finger motifs present in Ho are important for cleavage. The most unexpected feature of Ho, however, is its extreme inefficiency. Maximal cleavage occurs when Ho is present at a concentration of 1 molecule/3 base pairs of substrate DNA. Even under these conditions, complete digestion requires >2 h. This inefficiency results from two characteristics of Ho. First, Ho recycles slowly from cleaved product to new substrate, in part because the enzyme has an affinity for one end of its double strand break product. Second, high levels of cleavage in the in vitro reaction correlate with the appearance of large protein-DNA aggregates. At optimal Ho concentrations, these latter aggregates, referred to as "florettes," have an ordered structure consisting of a densely staining central region and loops of radiating DNA. These unusual properties may indicate that Ho plays a role in other aspects of mating type switching subsequent to double strand break formation.
Assuntos
DNA Fúngico/genética , Desoxirribonucleases de Sítio Específico do Tipo II/genética , Saccharomyces cerevisiae/genética , DNA Fúngico/química , Desoxirribonucleases de Sítio Específico do Tipo II/química , Plasmídeos/genética , Proteínas de Saccharomyces cerevisiaeRESUMO
The plastid genome in higher plants encodes subunits of an Escherichia coli-like RNA polymerase which initiates transcription of plastid genes from sequences resembling E.coli sigma70-type promoters. By deleting the gene for the essential beta subunit of the tobacco E.coli-like RNA polymerase, we have established the existence of a second plastid transcription system which does not utilize E.coli-like promoters. In contrast to the E.coli-like RNA polymerase, the novel transcription machinery preferentially transcribes genetic system genes rather than photosynthetic genes. Although the mutant plants are photosynthetically defective, transcription by this polymerase is sufficient for plastid maintenance and plant development.
Assuntos
RNA Polimerases Dirigidas por DNA/genética , Regulação da Expressão Gênica de Plantas , Nicotiana/genética , Plantas Tóxicas , Plastídeos/enzimologia , Sequência de Bases , Dados de Sequência Molecular , Fotossíntese , Regiões Promotoras Genéticas , RNA Ribossômico 16S/genética , Deleção de Sequência , Fator sigma/genética , Transcrição GênicaRESUMO
On infection of Escherichia coli cells by bacteriophages T4, T5 or T7, the degradation of E. coli protein fragments and abnormal proteins is inhibited. Normal E. coli proteins, however, continue to be degraded at their usual rates. T4 early proteins (s) is needed to inhibit the turnover of abnormal proteins in T4-infected E. coli cells.
Assuntos
Proteínas de Bactérias/metabolismo , Colífagos/fisiologia , Escherichia coli/metabolismo , Canavanina/metabolismo , Peptídeos/metabolismo , Proteínas Virais/metabolismoRESUMO
The cloned bacteriophage T4 pin gene functions to stabilize several different kinds of proteins in Escherichia coli bacteria. Incomplete proteins such as puromycyl polypeptides, abnormal but complete proteins such as the lambda phage tsO protein, and labile eukaryotic proteins encoded by genes cloned in E. coli such as mature human fibroblast interferon are stabilized in cells in which the T4 pin gene is expressed. The cloned T4 pin gene does not seem to affect the turnover of normal E. coli proteins.
Assuntos
Clonagem Molecular/métodos , Escherichia coli/genética , Genes Virais , Fagos T/genética , Proteínas de Bactérias/metabolismo , Interferon Tipo I/genética , Interferon Tipo I/metabolismoRESUMO
An Escherichia coli mutant, HDF026, defective for growth of phage T4, has been characterized biochemically and genetically. The mutant displays an elevated level of degradation of abnormal proteins, such as puromycyl polypeptides or canavanine-containing polypeptides. Genetically, HDF026 appears to be an allele of rho, which also encodes the transcription termination factor and RNA-dependent ATPase, Rho. The mutation contransduces by phage PI with ilv, weakly suppresses polar mutations in gal, and permits some growth of lambda N- phage. Temperature sensitive lambda mutants in gene O exhibit a reduced efficiency of plating at intermediate temperature on HDF026 mutants; presumably the lambda Ots protein is rapidly degraded in these strains. The ability of wild-type lambda to grow on HDF026 is also reduced, apparently the result of the lambda N product deficiency. gal escape synthesis, which reflects the level of lambda N activity, is decreased 50-66% in the HDF026 mutant. lambda r32, which requires more N function than wild-type phage, does not grow at all in HDF026. A lon mutation, which decreases protein degradation, partially reverses some of these phenotypes, suggesting that they are related to the protein hyperlability of HDF026.