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J Modolell

Publications and source records attributed to J Modolell.

At least 55 records · Page 3Linked to original sources
PubMed1987

Deletion analysis of the achaete-scute locus of Drosophila melanogaster.

The achaete-scute gene complex (AS-C) is involved in the development of the central and peripheral (sensory chaetae, sensilla) nervous system. To assess the contribution of the different parts of the complex in the generation of the adult chaetae pattern, we have determined the phenotypes and molecular positions of the breakpoints of 74 terminal deficiencies of the X chromosome. According to these and previous data, the AS-C is organized, distally to proximally, as follows: the achaete region, with most of its DNA (10 kb) located upstream from the putative achaete (T5) gene; an intermediate region, approximately 18 kb long, whose deletion only weakly affects the scute function; and the scute region, with most of the DNA critical for its function extending 4-5 kb upstream and 50 kb downstream of the putative scute (T4) gene. The DNA extending far upstream of the T5 gene and downstream of the T4 gene may provide chromatin conformations adequate for efficient expression of these genes. However, in the case of the T4 gene, the available data suggest the presence of a small number of elements, scattered in the long downstream region, that would respond to topological cues and cis-activate this gene in specific anatomical regions.

Animals↗
PubMed1986

Excess function hairy-wing mutations caused by gypsy and copia insertions within structural genes of the achaete-scute locus of Drosophila.

Hairy-wing (Hw) mutations cause the differentiation of extra chaetes on the cuticle of Drosophila. They are associated with modifications of the achaete-scute complex that consist, in the mutants studied, of insertions of the transposable elements gypsy (Hw1, HwBS) or copia (HwUa). gypsy and copia are inserted in achaete and scute transcribed regions, respectively. Transcription of the insertion-split genes starts at the normal site but terminates within the transposable element sequences. The RNA truncated within gypsy is 5-20 times more abundant than its homolog in wild-type flies. The abundance is reduced in Hw1 revertants and Hw1 stocks carrying su(Hw) mutations. These and other data suggest that the excess function phenotypes of Hw mutations are generated by an increase in achaete or scute transcripts.

Animals↗
PubMed1985

Molecular genetics of the achaete-scute gene complex of D. melanogaster.

The achaete-scute gene complex (AS-C), involved in differentiation of the sensory chaetes of D. melanogaster, and the yellow locus have been cloned. The yellow locus is the most distal and is followed, proximally, by the achaete and the scute loci. In the scute locus (75 kb), three transcription units separated by long stretches of DNA give rise to poly(A)+ RNAs of 1.6, 1.2, and 1.6 kb. Most DNA lesions associated with scute mutations map within the presumably untranscribed DNA. Their mutant phenotypes are stronger the closer the lesions are to the structural gene of one transcript (T4 RNA). Genetic and developmental data suggest that only this RNA is fundamental for the scute function. Its transcription might be perturbed by far removed DNA lesions. A second transcript is probably implicated in the lethal of scute embryonic function, while the third transcript is unnecessary for the differentiation of most macrochaetes. Two additional polyadenylated RNAs are transcribed from the achaete (1.1 kb) and yellow (1.9 kb) loci.

Animals↗
PubMed1983

Drosophila melanogaster mutations suppressible by the suppressor of Hairy-wing are insertions of a 7.3-kilobase mobile element.

Certain spontaneous mutations of Drosophila melanogaster are suppressed by su(Hw), the suppressor of Hairy-wing (3R-54.8). We find that mutations suppressible by su(Hw) result from insertions of a mobile element at the affected loci. The element, named gypsy, is approximately 7.3 kilobases long and includes 0.5-kilobase direct terminal repeats. It was first identified in DNA cloned from the bithorax chromosomal region of several Drosophila stocks carrying suppressible mutations of the bithorax complex. Cloned gypsy DNA was used as a probe to test for the association of gypsy with suppressible mutations at various other loci by hybridization in situ. Gypsy was found to be associated with 19 suppressible alleles at 10 different loci: yellow, Hairy-wing, scute, diminutive, cut, lozenge, forked, Beadex, hairy, and the bithorax complex. It was found with wild-type or nonsuppressible mutations at any of these loci. Gypsy DNA was also used as a probe to clone the element and adjacent unique DNA from the loci of some suppressible mutations. This confirmed the presence of the full-length element and also provided cloned DNA from the previously uncloned loci scute and cut. The suppressor of Hairy-wing is generally recessive and behaves as a null mutation. Thus, the disruption of normal gene function caused by the inserted gypsy element appears to require some product of the wild-type suppressor gene, su(Hw)+.

Animals↗
PubMed1982

DNA map of mutations at the scute locus of Drosophila melanogaster.

The achaete-scute gene complex (AS-C) of Drosophila melanogaster is involved in the differentiation of innervated elements in the adult (chaetes) and in the embryo (central nervous system). Genetically, the AS-C is subdivided into four regions: achaete, scute alpha, lethal of scute, and scute beta. Using a previously cloned fragment of scute DNA, we have now cloned 62 kb of wild-type DNA from the scute region. No repetitive sequences have been detected in this stretch of DNA. Of 16 scute mutants with chromosomal rearrangements studied (inversions, deletions, and translocations), nine, included genetically in scute beta, have breakpoints in the cloned region. The remaining rearrangements, which genetically correspond to scute alpha, map outside and to the left of the cloned region. Of nine scute ;point mutants' studied, eight have large DNA alterations within the cloned region. These alterations include insertions (five) and deletions (three). The DNA alterations found in both ;point mutants' and rearrangements are interspersed and scattered over 40 kb. The relationship between the sites of the DNA alterations and the mutant phenotypes are discussed.

Journal Article↗
PubMed1981

Effects of antibiotics, N-acetylaminoacyl-tRNA and other agents on the elongation-factor-Tu dependent and ribosome-dependent GTP hydrolysis promoted by 2'(3')-O-L-phenylalanyladenosine.

GTP hydrolysis on elongation factor (EF) Tu . ribosome complexes has been assayed in the presence of 2'(3')-O-L-phenylalanyladenosine (AdoPhe), i.e. the 3'-terminal portion of Phe-tRNAPhe. Several requirements of the reaction have been characterized. Maximal activity is observed at 60-120 mM NH4Cl and 5-15 mM magnesium acetate. The reaction requires the free sulfhydryl group of EF-Tu normally implicated in aminoacyl-tRNA binding. Intact EF-Tu cannot be replaced by a large tryptic fragment of EF-Tu (Mr 39,000) that retains the ability to bind guanosine nucleotides. The aminoglycoside antibiotics, neomycin C and several kanamycins and gentamicins, stimulate the AdoPhe-promoted GTPase. Surprisingly, however, other closely related antibiotics, like neomycin B, paromomycin and ribostamycin, are ineffectual, thus indicating subtle differences in the actions of these antibiotics. AcPhe-tRNAPhe, bound to the ribosomal A-site, stimulates the AdoPhe-promoted GTPase, but this compound or AcTyr-tRNATyr, present in unbound form, strongly inhibits the reaction. These results suggest that N-blocked aminoacyl-tRNAs form ternary complexes with EF-Tu . GTP, which have not been previously detected because of their low stability.

Anti-Bacterial Agents↗
PubMed1980

Nonenzymic translocation and spontaneous release of noncognate peptidyl transfer ribonucleic acid from Escherichia coli ribosomes.

Poly(uridylic acid)-programmed ribosomes have been used to synthesize the noncognate peptidyl-tRNA Ac-Phe-Tyr-tRNATyr and its cognate counterpart Ac(Phe)2-tRNAPhe. After synthesis, Ac(Phe)2-tRNAPhe remains, as expected, in the ribosomal acceptor (A) site, but the noncognate AcPhe-Tyr-tRNATyr does not; part of it spontaneously falls off the ribosome and the rest translocates, without elongation factor (EF) G, to the ribosomal donor site. The inhibitor of translocation viomycin prevents both the spontaneous release and the nonenzymatic translocation by confining the noncognate peptidyl-tRNA to the A site. Under these conditions, the interaction of AcPhe-Tyr-tRNATyr with the A site appears to be similar to that of Ac(Phe)2-tRNAPhe without the antibiotic, and EF-G promotes the translocation and subsequent elongation of both peptidyl-tRNAs to comparable extents. The results indicate that, without viomycin, the noncognate peptidyl-tRNA is weakly held in the ribosomal A site and support the proposal that the release of peptidyl-tRNA occurring during protein synthesis in vivo is related to a ribosomal editing mechanism which discards mistranslated nascent proteins [Menninger, J. R. (1977) Mech. Ageing Dev. 6, 131].

Escherichia coli↗
PubMed1980

Hydrolysis of GTP on elongation factor Tu.ribosome complexes promoted by 2'(3')-O-L-phenylalanyladenosine.

In the presence of Escherichia coli ribosomes and elongation factor EF) Tu, 2'(3')-O-L-phenylalanyladenosine (AdoPhe), the 3'-terminal portion of Phe-tRNAPhe, promotes the hydrolysis of GTP. The reaction requires the presence of both 30S and 50S ribosomal subunits and of proteins L7/L12 on the 50S subunit, is unaffected by mRNA [poly(uridylic acid)], and is strongly stimulated by EF-Ts. It is proposed that the AdoPhe-dependent GTP hydrolysis, like that promoted by aminoacyl-tRNA, is mediated by a ternary complex with EF-Tu and GTP; however, in contrast to aminoacyl-tRNA, AdoPhe is probably not retained by ribosomes after GTP hydrolysis. Phe-tRNAPhe or N-acetyl-Phe-tRNAPhe bound to the ribosomal acceptor site do not inhibit, but even stimulate, GTP hydrolysis by AdoPhe.EF-Tu.GTP. Thus, the binding site for EF-Tu on the ribosome is probably available for interaction with AdoPhe.EF-Tu.GTP regardless of whether the nearby acceptor site is vacant of occupied with aminoacyl-tRNA or peptidyl-tRNA. The results demonstrate the critical role of the 3'-terminal region of aminoacyl-tRNA in activating the EF-Tu- plus ribosome-dependent GTPase.

Adenosine↗
PubMed1980

Hygromycin A, a novel inhibitor of ribosomal peptidyltransferase.

In cell-free systems from Escherichia coli, hygromycin A inhibits polypeptide synthesis directed by either poly(U) or phage R 17 RNA, and the reaction of puromycin with either natural peptidyl-tRNA, or AcPhe-tRNA, or the 3'-terminal fragment of AcLeu-tRNA (C-A-C-C-A-LeuAc). In contrast, the antibiotic does no inhibit the enzymatic binding of Phe-tRNA to ribosomes or the translocation of AcPhe-tRNA. It is concluded that hygromycin A is a specific inhibitor of the peptide bond formation step of protein synthesis. The action of hygromycin A on peptidyl transfer is similar to that of chloramphenicol, an antibiotic that shares some common structural features with hygromycin A. Both antibiotics inhibit the binding of C-A-C-C-A-Leu to the acceptor site of peptidyl transferase and stimulate that of C-A-C-C-A-LeuAc to the donor site of the enzyme. Moreover, hygromycin A blocks the binding of chloramphenicol to ribosomes, indicating that the binding sites of the antibiotics may be closely related. Hygromycin A is a more potent agent than chloramphenicol and binds quite strongly to ribosomes.

Acyltransferases↗
PubMed1979

Elongation factor Tu-induced conformational changes of ribosomes detected by iodination.

The effect of elongation factor (EF) Tu, bound to the ribosome with the help of poly(uridylic) acid, Phe-tRNA and guanyl-5'-yl methylene diphosphonate, on the conformation and/or chemical environment of ribosomal proteins has been examined using, as a probe, protein iodination. Ribosomes complexed only with poly(uridylic acid) and Phe-tRNA have been used as a control. EF-Tu on the ribosome significantly increases the iodination of proteins S7, S10 and L3 and decreases that of S21 and L18.

Lactoperoxidase↗
PubMed1979

Dissociation of guanosine nucleotide-elongation factor G-ribosome complexes.

The spontaneous dissociation of complexes containing elongation factor G (EF-G), the ribosome, and either GDP plus fusidic acid, guanyl-5'-yl imidodiphosphate, or guanyl-5'-yl methylene diphosphonate has been measured and it follows biphasic kinetics that can be resolved into two first-order decay rates. This suggest the existence of two classes of complexes with apparent dissociation rate constants (k) differing 5--20-fold. The values of k and the distribution of complexes between the fast and the slowly decaying class depend on the conditions in which the dissociation occurs but not on the conditions in which the complexes are formed. Rapid transitions of complexes from one to the other class occur only when the chemical environment in which the dissociation takes place is modified. Thus, increasing the concentration of NH4Cl or adding the antibiotic thiostrepton accelerates the decay and converts slowly dissociating into fast dissociating complexes. In contrast, addition of misreading-inducing aminoglycoside antibiotics of the neomycin, kanamycin, streptomycin, and gentamicin (but not hygromycin) groups slows down the decay. For neomycin B at 10 micron, this effect is due to the conversion of fast into slowly decaying complexes. A model to explain the results involving conformational transitions of the complexes is proposed.

Anti-Bacterial Agents↗
PubMed1979

The binding of non-cognate Tyr-tRNATyr to poly(uridylic acid)-programmed Escherichia coli ribosomes.

The poly(U)-dependent binding of Tyr-tRNATyr to Escherichia coli ribosomes has been studied using a highly purified system. Binding is maximal at 10 mM magnesium acetate (up to 0.7 molecule Tyr-tRNATyr/ribosome), and requires the presence of elongation factor (EF) T (a mixture of EF-Ts and EF-Tu), GTP, NH4+ ions and an aminoglycoside antibiotic (streptomycin, neomycin B, kanamycin B or gentamicin C1a). Under limiting and up to saturating concentrations of EF-T, one molecule of GTP is hydrolyzed per molecule of Tyr-tRNATyr bound, suggesting that 'proof-reading' mechanisms involving the hydrolysis of GTP are inoperative in the presence of the antibiotics. Binding of Tyr-tRNATyr apparently takes place at the ribosomal acceptor site, since peptide bonds are readily formed with N-acetyl-Phe-tRNA prebound to the ribosomal donor site. In contrast to Phe-tRNAPhe binding, Tyr-tRNATyr binding is impaired by the omission of the 50-S subunit, the replacement of GTP by its non-hydrolyzable analogs guanyl-5'-yl methylene diphosphonate and guanyl-5'-yl iminodiphosphonate, and also by the presence of the antibiotic streptogramin A. This suggests that the correct interaction of Tyr-tRNATyr with the peptidyl transferase centre is essential for the stability of this ligand on the ribosome. Moreover, the aminoglycoside antibiotics are also necessary, even after the binding reaction is complete, to maintain Tyr-tRNATyr on the ribosome.

Aminoglycosides↗