Primary Sequence of the \u3ci\u3eEscherichia coli fadBA\u3c/i\u3e Operon, Encoding the Fatty Acid-Oxidizing Multienzyme Complex, Indicates a High Degree of Homology to Eucaryotic Enzymes

In Escherichia coli at least five enzyme activities required for the beta-oxidation of fatty acids are associated with a multienzyme complex composed of two subunits in α2β2 conformation (A. Pramanik et al., J. Bacteriol. 137:469-473, 1979). In the present work, the DNA sequence of the genes encoding these two subunits, fadB and fadA, has been determined. The direction of transcription was from fadB to fadA rather than from fadA to fadB, as suggested previously (S. K. Spratt et al., J. Bacteriol. 158:535-542, 1984). Only 10 nucleotides separated the coding sequences for the two peptides, confirming the suggestion that these genes form an operon. The peptides encoded by fadB and fadA were 729 amino acids and 387 amino acids, respectively, in length. The larger and smaller peptides had predicted molecular masses of 79,678 and 40,876 Da, respectively. Recently, the sequence of thefadA gene was published in a separate report (Yang et al., J. Biol. Chem. 265:10424-10429, 1990). In this work, most of the DNA sequence for fad4 was confirmed, and 10 errors were corrected. Three of these nucleotide changes resulted in five amino acid residue changes predicted in the carboxy terminus of the fadA-encoded peptide. By comparison to other peptide sequences, the a subunit encoded withinfadB had 31% perfect identity with the rat peroxisomal enoyl-coenzyme A:hydratase-3-hydroxyacyl-coenzyme A dehydrogenase trifunctional enzyme over the entire length of the two peptides. In agreement with the work of Yang et al., the β subunit encoded within fadA had 35 to 45% perfect identity with five thiolase genes from different eucaryotic sources over the entire length of the peptide

Primary Sequence of the \u3ci\u3eEscherichia coli fadBA\u3c/i\u3e Operon, Encoding the Fatty Acid-Oxidizing Multienzyme Complex, Indicates a High Degree of Homology to Eucaryotic Enzymes

In Escherichia coli at least five enzyme activities required for the beta-oxidation of fatty acids are associated with a multienzyme complex composed of two subunits in α2β2 conformation (A. Pramanik et al., J. Bacteriol. 137:469-473, 1979). In the present work, the DNA sequence of the genes encoding these two subunits, fadB and fadA, has been determined. The direction of transcription was from fadB to fadA rather than from fadA to fadB, as suggested previously (S. K. Spratt et al., J. Bacteriol. 158:535-542, 1984). Only 10 nucleotides separated the coding sequences for the two peptides, confirming the suggestion that these genes form an operon. The peptides encoded by fadB and fadA were 729 amino acids and 387 amino acids, respectively, in length. The larger and smaller peptides had predicted molecular masses of 79,678 and 40,876 Da, respectively. Recently, the sequence of thefadA gene was published in a separate report (Yang et al., J. Biol. Chem. 265:10424-10429, 1990). In this work, most of the DNA sequence for fad4 was confirmed, and 10 errors were corrected. Three of these nucleotide changes resulted in five amino acid residue changes predicted in the carboxy terminus of the fadA-encoded peptide. By comparison to other peptide sequences, the a subunit encoded withinfadB had 31% perfect identity with the rat peroxisomal enoyl-coenzyme A:hydratase-3-hydroxyacyl-coenzyme A dehydrogenase trifunctional enzyme over the entire length of the two peptides. In agreement with the work of Yang et al., the β subunit encoded within fadA had 35 to 45% perfect identity with five thiolase genes from different eucaryotic sources over the entire length of the peptide

Nucleotide sequence of the \u3ci\u3efadR\u3c/i\u3e gene, a multifunctional regulator of fatty acid metabolism in \u3ci\u3eEscherichia coli\u3c/i\u3e

The Escherichia coli fadR gene is a multifunctional regulator of fatty acid and acetate metabolism. In the present work the nucleotide sequence of the 1.3 kb DNA fragment which encodes FadR has been determined. The coding sequence of the fadR gene is 714 nucleotides long and is preceded by a typical E. coli ribosome binding site and is followed by a sequence predicted to be sufficient for factor-independent chain termination. Primer extension experiments demonstrated that the transcription of the fadR gene initiates with an adenine nucleotide 33 nucleotides upstream from the predicted start of translation. The derived fadR peptide has a calculated molecular weight of 26,972. This is in reasonable agreement with the apparent molecular weight of 29,000 previously estimated on the basis of maxi-cell analysis of plasmid encoded proteins. There is a segment of twenty amino acids within the predicted peptide which resembles the DNA recognition and binding site of many transcriptional regulatory proteins

cis-acting sequences and trans-acting factors in the localization of mRNA for mitochondrial ribosomal proteins

mRNA localization is a conserved post-transcriptional process crucial for a variety of systems. Although several mechanisms have been identified, emerging evidence suggests that most transcripts reach the protein functional site by moving along cytoskeleton elements. We demonstrated previously that mRNA for mitochondrial ribosomal proteins are asymmetrically distributed in the cytoplasm, and that localization in the proximity of mitochondria is mediated by the 3′-UTR. Here we show by biochemical analysis that these mRNA transcripts are associated with the cytoskeleton through the microtubule network. Cytoskeleton association is functional for their intracellular localization near the mitochondrion, and the 3′-UTR is involved in this cytoskeleton-dependent localization. To identify the minimal elements required for localization, we generated DNA constructs containing, downstream from the GFP gene, deletion mutants of mitochondrial ribosomal protein S12 3′-UTR, and expressed them in HeLa cells. RT-PCR analysis showed that the localization signals responsible for mRNA localization are located in the first 154 nucleotides. RNA pulldown assays, mass spectrometry, and RNP immunoprecipitation assay experiments, demonstrated that mitochondrial ribosomal protein S12 3′-UTR interacts specifically with TRAP1 (tumor necrosis factor receptor-associated protein1), hnRNPM4 (heterogeneous nuclear ribonucleoprotein M4), Hsp70 and Hsp60 (heat shock proteins 70 and 60), and α-tubulin in vitro and in vivo

COMPOUNDS FOR INCREASING LIPID SYNTHESIS AND STORAGE

This invention relates to methods for increasing lipid production in cells. Methods of producing biofuel from cells and preparing mutraceuticals comprising lipids produced according to a method provided herein are also provided

Compression of time during smooth pursuit eye movements

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Cloning and Characterization of a Gene (fadR) Involved in Regulation of Fatty Acid Metabolism in \u3ci\u3eEscherichia coli\u3c/i\u3e

The regulatory gene fadR has been previously characterized by classical genetic means as a diffusible protein which exerts negative control over fatty acid degradation and acetate metabolism. fadR has also been implicated in the regulation of unsaturated fatty acid biosynthesis. To facilitate the identification of the product of the fadR gene and to study the mechanism by which this multifunctional regulatory gene exerts its control, we cloned a segment of DNA containing the fadR gene in the phage vector λL47. Subsequent subcloning of a segment of the chromosomal DNA from the λfadR+ phage into various plasmid vectors resulted in the isolation of the fadR gene on a 1.3-kilobase-pair HindIII-EcoRV fragment. fadR strains harboring the clonedfadR\u27 gene showed inducible levels of fatty acid oxidation and crotonase (enoyl-coenzyme A-hydratase, fadB) activity. The cloned gene exerted transcriptional control over 13-galactosidase synthesis in an fadR strain that had a λΦ(fadE-lacZ+) operon fusion. An fadR mutation in fabA(Ts) strains prevents growth at permissive temperatures without unsaturated fatty acid supplementation (Nunn et al., J. Bacteriol. 154:554-560, 1983). Plasmids carrying the fadR+ gene suppress this unsaturated fatty acid auxotrophy in fadR fabA(Ts) strains at the permissive condition. Maxiceli analysis identified a 29,000-dalton protein encoded by the 1.3-kilobase fragment which appeared to be associated with functional fadR gene activity

Cloning and Characterization of a Gene (fadR) Involved in Regulation of Fatty Acid Metabolism in \u3ci\u3eEscherichia coli\u3c/i\u3e

The regulatory gene fadR has been previously characterized by classical genetic means as a diffusible protein which exerts negative control over fatty acid degradation and acetate metabolism. fadR has also been implicated in the regulation of unsaturated fatty acid biosynthesis. To facilitate the identification of the product of the fadR gene and to study the mechanism by which this multifunctional regulatory gene exerts its control, we cloned a segment of DNA containing the fadR gene in the phage vector λL47. Subsequent subcloning of a segment of the chromosomal DNA from the λfadR+ phage into various plasmid vectors resulted in the isolation of the fadR gene on a 1.3-kilobase-pair HindIII-EcoRV fragment. fadR strains harboring the clonedfadR\u27 gene showed inducible levels of fatty acid oxidation and crotonase (enoyl-coenzyme A-hydratase, fadB) activity. The cloned gene exerted transcriptional control over 13-galactosidase synthesis in an fadR strain that had a λΦ(fadE-lacZ+) operon fusion. An fadR mutation in fabA(Ts) strains prevents growth at permissive temperatures without unsaturated fatty acid supplementation (Nunn et al., J. Bacteriol. 154:554-560, 1983). Plasmids carrying the fadR+ gene suppress this unsaturated fatty acid auxotrophy in fadR fabA(Ts) strains at the permissive condition. Maxiceli analysis identified a 29,000-dalton protein encoded by the 1.3-kilobase fragment which appeared to be associated with functional fadR gene activity

Income and childbearing decisions: evidence from Italy

During the early 1990s, Italy has been one of the first countries to reach lowest-low fertility, i.e. below 1.3 children per woman. In this paper we focus on the period during which such fertility levels arose in order to assess the impact of income on fertility decisions. So far, analyses have suffered from the lack of appropriate data; we create a new data set making use of two different surveys from Bank of Italy (SHIW) and ISTAT (Labor Force Survey) and we apply discrete-time duration models. For first births, we find evidence of non-proportional hazards and of some recuperation effect: women with high predicted wages tend to delay the first birth, subsequently recuperating. For second and third births, instead, the availability of a good child-care system seems to play a key role and income exhibits small intensity. In a final section, we explore the possible effect on fertility of an increase in financial support for poorer families that took place in 1999

Mirabilia Coralii. Capolavori barocchi in corallo tra maestranze ebraiche e trapanesi

La pregevole lavorazione del corallo trapanese ricostruita attraverso la puntuale analisi delle fonti documentarie, manoscritte e bibliografiche. L’organizzazione della maestranza e il ruolo che la committenza,sia laica che ecclesiastica, ha esercitato nella realizzazione di capolavori oggi presenti in rinomate collezioni pubbliche e private e nei principali musei nazionali ed internazionali. Una rassegna di importanti manufatti, un tempo mirabili opere da Wunderkammer, criticamente commentata, che bene evidenzia l’aulica produzione siciliana ricercata da Sovrani, Viceré ed alti prelati. Un’originale ed inedita trattazione che per la prima volta rende noti nomi e biografie di gran parte degli artisti specializzati, tra XVI e XIX secolo, nella trasformazione del prezioso materiale marin