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G Ditta

Publications and source records attributed to G Ditta.

15 recordsLinked to original sources
PubMed1997

Floral determination and expression of floral regulatory genes in Arabidopsis.

The expression of the floral regulators LEAFY, APETALA1 and AGAMOUS-LIKE8 was examined during light treatments that induced flowering in Arabidopsis, and was compared to time points at which floral determination occurred. Extension of an 8-hour day by either continuous red- or far-red-enriched light induced LEAFY and AGAMOUS-LIKE8 expression within 4 hours. The 4 hours of additional light was sufficient for floral determination only in the far-red-enriched conditions, while 12-16 hours of additional light was required for floral determination in the red-enriched conditions. These results indicate that the induction of floral regulatory genes and induction of flower formation can be uncoupled under certain circumstances. Expression of LEAFY and AGAMOUS-LIKE8 in the shoot apex at the time of floral determination is also consistent with genetic data indicating that these genes are involved in the first steps of the transition from vegetative to reproductive development. In contrast to LEAFY and AGAMOUS-LIKE8, APETALA1 expression was first observed 16 hours after the start of photoinduction. Since this time point was always after floral determination, APETALA1 is an indicator of floral determination.

Arabidopsis↗
PubMed1991

Modular structure of FixJ: homology of the transcriptional activator domain with the -35 binding domain of sigma factors.

The FixL/FixJ two-component system is a global regulator of nitrogen-fixation genes in Rhizobium meliloti. The transcriptional activator FixJ contains two modules: its N-terminal module is homologous with other two-component regulators; its C-terminal module shows homology with various transcriptional activators, and with the C-terminal region of sigma factors, which is involved in the discrimination of the -35 region of bacterial promoters. We show that the C-terminal module of FixJ contains the entire transcription activation function, and that the N-terminal module regulates this activity negatively. Oligonucleotide-directed mutagenesis of the transcriptional activator module demonstrated the importance of a potential helix-turn-helix structure.

Amino Acid Sequence↗
PubMed1990

Differential expression of hydrogen uptake (hup) genes in vegetative and symbiotic cells of Rhizobium leguminosarum.

The genetic determinants responsible for H2-uptake (hup genes) in Rhizobium leguminosarum are organized in six transcriptional units, designated regions hupI to hupVI, with region hupI coding for the hydrogenase structural genes (Leyva et al. 1990). Regulation of the expression of hup genes from R. leguminosarum was examined by using hup-lacZ fusions and mRNA dot-blot analysis. None of the six hup regions is transcribed in vegetative cells grown under normal aerobic conditions, whereas all six regions are transcribed in pea bacteroids. Additionally, exposure of cell cultures to low oxygen tensions specifically induces the expression of regions hupV and hupVI. By studying the expression of hupV- and hupVI-lacZ fusions in R. meliloti mutants it was determined that the microaerobic induction of these two regions is dependent on the regulatory fixLJ system, and that this control is exerted through fixK. Such expression was also shown to be nifA and ntrA independent. The functions of the hupV and hupVI gene products are unknown. The possibility that they play a regulatory role in hup gene expression is unlikely, since pea bacteroids from R. leguminosarum Hup- mutants carrying Tn5 insertions in regions hupV and hupVI contained normal levels of mRNA transcripts corresponding to the remaining hup regions.

DNA Transposable Elements↗
PubMed1987

The nifA gene of Rhizobium meliloti is oxygen regulated.

Experiments using plasmid-borne gene fusions and direct RNA measurements have revealed that expression from the nifA gene is induced in Rhizobium meliloti when the external oxygen concentration is reduced to microaerobic levels. Induction occurs in the absence of alfalfa and in the presence of fixed nitrogen and does not require ntrC. The production of functional nifA gene product (NifA) can be demonstrated by its ability to activate the nitrogenase promoter P1. Aerobic induction of nifA can also occur during nitrogen starvation at low pH, but in this case induction is dependent on ntrC and does not lead to P1 activation. The data indicate that reduced oxygen tension is potentially a major trigger for symbiotic activation of nitrogen fixation in Rhizobium species.

Energy Metabolism↗
PubMed1986

Overlapping transcription of the nifA regulatory gene in Rhizobium meliloti.

Gene fusions were used to demonstrate complex overlapping transcriptional controls of the regulatory gene nifA in Rhizobium meliloti bacteroids isolated from alfalfa root nodules. Gene nifA has previously been shown to be transcribed from promoter PnifA and to be required for activation of promoters P1 (nifHDK) and P2 (fixABC) during symbiotic nitrogen fixation with alfalfa. P2 is located approximately 4 kb upstream from nifA and is shown in this report to be responsible for at least one-half of the nifA expression observed in bacteroids. Substantial transcription of nifA occurs during symbiosis as evidenced by the fact that PnifA was found to be 53% as active as either P1 or P2. Together, the data indicate that more than half of the transcripts initiated at P2 fail to terminate before reaching nifA. Additional studies indicated that there may be weak promoter activity in the symbiotically essential region downstream from nifA.

Acetyltransferases↗
PubMed1986

Rhizobium meliloti genes required for nodule development are related to chromosomal virulence genes in Agrobacterium tumefaciens.

Symbiotically essential genes have been identified in Rhizobium meliloti that are structurally and functionally related to chromosomal virulence (chv) genes of Agrobacterium tumefaciens. Homologous sequences also exist in the genomes of other fast-growing rhizobia including Rhizobium trifolii, Rhizobium leguminosarum, and Rhizobium phaseoli. In Agrobacterium, the chvA and chvB loci are known to be essential for oncogenic transformation of dicotyledonous plants and for attachment to plant cells [Douglas, C. J., Staneloni, R. J., Rubin, R. A. & Nester, E. W. (1985) J. Bacteriol. 64, 102-106], and the chvB locus has been implicated in the production of (1-->2)-beta-glucan, a unique exopolysaccharide component [Puvanesarajah, V., Schell, F. M., Stacey, G., Douglas, C. J. & Nester, E. W. (1985) J. Bacteriol. 164, 102-106]. Site-directed transposon insertion mutants in the chvA and chvB-equivalent regions of R. meliloti are symbiotically defective. Mutants in the chvB-equivalent region have been examined in detail and have been found to induce the formation of nodule-like structures on alfalfa that are devoid of bacteroids, lack infection threads, and cannot fix nitrogen. Such mutants fluoresce normally in the presence of Calcofluor, a histochemical stain for beta-linked polysaccharides, and produce normal amounts of total exopolysaccharide. The Rhizobium loci have been designated ndv because of their requirement for nodule development.

Journal Article↗
PubMed1985

Deletion analysis of Rhizobium meliloti symbiotic promoters.

Previous examination of DNA sequences located 5' to Rhizobium meliloti nif transcription units has shown that extensive sequence homology exists among them. Here we have examined these reiterated sequences for their role in symbiotic gene regulation. Promoter deletion analysis has shown that although an extensive upstream DNA sequence 9160 bp) is required for full heterologous activation of the R. meliloti nifHDK promoter by the Klebsiella pneumoniae nifA protein in Escherichia coli, this region is not required for expression of R. Meliloti nif promoters in root nodules from plants grown under greenhouse conditions. In addition, a minimum functional symbiotic promoter sequence is defined, and a DNA sequence difference affecting regulation of two symbiotic promoters is discussed.

Bacterial Proteins↗
PubMed1983

Structural relationships among Rhizobium meliloti symbiotic promoters.

Symbiotic nitrogen fixation by Rhizobium meliloti requires the developmentally specific expression of certain bacterial genes. One set of these genes encodes the subunits of nitrogenase, the enzyme responsible for the reduction of atmospheric dinitrogen to ammonia, and another set consists of closely linked genes also essential for nitrogen fixation. Examination of promoter and probable regulatory regions for these gene sets has revealed extensive DNA sequence conservation for more than 160 bp upstream of the respective transcription start points. Three such promoter regions have been identified in the nitrogen fixation (nif) gene cluster of R. meliloti strain 102F34. Using one of these promoter regions as a hybridization probe, three additional sequences were found in the genome of this strain. The DNA of other R. meliloti strains and Rhizobium species were also examined for homology to the symbiotically regulated promoters of R. meliloti 102F34. DNA sequences homologous to these R. meliloti promoters were found among diverse rhizobia, and in at least some cases were associated with nif genes.

Base Sequence↗
PubMed1983

Organization and expression of Rhizobium meliloti nitrogen fixation genes.

The boundaries of a nif gene cluster in Rhizobium meliloti were determined by Tn5 mutagenesis. These genes are clustered within a 14- to 15-kilobase (kb) region that includes the nitrogenase structural genes. Sequences within 10 kb on either side of this region are devoid of symbiotically essential gene function. RNA blot analysis identified a 5- to 6-kb transcript that corresponds to the nitrogenase structural gene operon. The 5' end of this transcript and its polarity were determined by nuclease S1 mapping. The 5' end of another nif transcript was also identified by nuclease S1 mapping. The promoter regions for these two nif transcripts control transcription in divergent directions and are separated by 1.9 kb of symbiotically unessential DNA. One Tn5 insertion within the nitrogenase operon did not create a polar mutation. The implications of this finding and the overall emerging picture of the genetic organization of this nif region are discussed.

Journal Article↗
PubMed1982

Clustering of nitrogen fixation (nif) genes in Rhizobium meliloti.

A cloned 17.3-kilobase region of the Rhizobium meliloti genome with homology to the Klebsiella pneumoniae nitrogenase structural genes was studied. Limits on the extent of homology were determined. Transposon mutagenesis of this region of the genome verified that it contained functional nif genes, Some transposon insertions resulted in a defective symbiotic phenotype, whereas others had no noticeable effect on symbiotic competence. The relative position of insertions yielding these two phenotypic classes suggested that at least three distinct units of gene expression are present in this region. Hybridization of RNA from alfalfa root nodules and from vegetatively grown Rhizobium to this cloned DNA showed that at least 11.1 kilobases of the region was transcribed actively and that transcription was specific for the symbiotic state.

Cloning, Molecular↗
PubMed1980

Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti.

A broad host range cloning vehicle that can be mobilized at high frequency into Gram-negative bacteria has been constructed from the naturally occurring antibiotic resistance plasmid RK2. The vehicle is 20 kilobase pairs in size, encodes tetracycline resistance, and contains two single restriction enzyme sites suitable for cloning. Mobilization is effected by a helper plasmid consisting of the RK2 transfer genes linked to a ColE1 replicon. By use of this plasmid vehicle, a gene bank of the DNA from a wild-type strain of Rhizobium meliloti has been constructed and established in Escherichia coli. One of the hybrid plasmids in the bank contains a DNA insert of approximately 26 kilobase pairs which has homology to the nitrogenase structural gene region of Klebsiella pneumoniae.

Cloning, Molecular↗
PubMed1976

The selection of Chinese hamster cells deficient in oxidative energy metabolism.

A selection scheme based on the nutritional requirements of a previously described respiration-deficient Chinese hamster line has been used to isolate new mutants defective in oxidative energy metabolism. Three of the primary characteristics of this type of mutant are (1) a strict dependency on the continued presence of glucose for survival; (2) a drastic reduction in the rate of oxygen consumption; (3) an inhibition of Krebs cycle activity resulting in auxotrophy for asparagine and carbon dioxide. In the case of one cell line which was used (V79), up to 65% of the survivors of a selection were found to possess this phenotype after only one round of selection. By contrast, it proved much more difficult to obtain such mutants from another cell line (CCL16). A preliminary characterization of a number of these mutants is presented.

Animals↗