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1.
Cell ; 93(5): 779-89, 1998 May 29.
Article in English | MEDLINE | ID: mdl-9630222

ABSTRACT

The organs of a higher plant show two fundamental axes of asymmetry: proximodistal and dorsoventral. Dorsoventrality in leaves, bracts, and petal lobes of Antirrhinum majus requires activity of the PHANTASTICA (PHAN) gene. Conditional mutants revealed that PHAN is also required for earlier elaboration of the proximodistal axis. PHAN was isolated and shown to encode a MYB transcription factor homolog. PHAN mRNA is first detected in organ initials before primordium initiation. The structure and expression pattern of PHAN, together with its requirement in two key features of organ development, are consistent with a role in specifying lateral organ identity as distinct from that of the stem or meristem. PHAN also appears to maintain meristem activity in a non-cell-autonomous manner.


Subject(s)
DNA-Binding Proteins/genetics , Genes, Plant , Magnoliopsida/growth & development , Plant Proteins/genetics , Plant Shoots/growth & development , Proto-Oncogene Proteins c-myb , Transcription Factors/genetics , Alleles , Amino Acid Sequence , Base Sequence , Cloning, Molecular , In Situ Hybridization , Magnoliopsida/genetics , Meristem/genetics , Meristem/growth & development , Molecular Sequence Data , Phenotype , Plant Shoots/genetics , RNA, Messenger/isolation & purification , RNA, Plant/isolation & purification , Sequence Homology, Amino Acid , Tissue Distribution
2.
J Bacteriol ; 175(14): 4405-13, 1993 Jul.
Article in English | MEDLINE | ID: mdl-8331071

ABSTRACT

The pcnB gene product of Escherchia coli is required for copy number maintenance of plasmids related to ColE1 and also for that of the IncFII plasmid R1. Because PcnB is similar to the tRNA-binding protein tRNA nucleotidyltransferase, we have suggested that the protein would be required only for processes in which an RNA is a prominent regulatory component. This appears to be so; strains deleted for pcnB, although defective in ColE1 and R1 plasmid maintenance, maintain the iteron-regulated plasmids F and P1 normally. We also find that strains deleted for pcnB grow normally, demonstrating that PcnB has no essential cellular role under the conditions tested and suggesting that regulation by antisense RNAs similar to RNAI has no critical role in any essential host process. We confirm by immunological tests that PcnB is likely to be the commercially available enzyme poly(A) polymerase.


Subject(s)
Bacterial Proteins/genetics , Escherichia coli Proteins , Escherichia coli/genetics , Genes, Bacterial , Plasmids , R Factors , Ampicillin/pharmacology , Ampicillin Resistance/genetics , Antibodies/metabolism , Bacterial Proteins/biosynthesis , Bacterial Proteins/isolation & purification , Chromosomes, Bacterial , DNA, Bacterial/metabolism , Escherichia coli/drug effects , Escherichia coli/metabolism , F Factor , Gene Deletion , Genotype , Glutathione Transferase/biosynthesis , Glutathione Transferase/genetics , Glutathione Transferase/isolation & purification , Polynucleotide Adenylyltransferase/metabolism , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/isolation & purification , Restriction Mapping
3.
J Bacteriol ; 174(7): 2102-10, 1992 Apr.
Article in English | MEDLINE | ID: mdl-1551833

ABSTRACT

The replication terminus region (31 to 35 min) of the Escherichia coli chromosome contains very few mapped genes (two per min) compared with the remainder of the chromosome, and much of the DNA appears dispensable. In order to determine whether, despite this, the terminus region consists of protein-coding sequences, we cloned 44 kb (1 min) of terminus region DNA (that surrounding trg at 31.4 min) and examined its ability to catalyze protein synthesis in vitro or in minicells. We were able to account for more than half the coding capacity of the cloned DNA with proteins synthesized in these systems, indicating that the sparsity of mapped genes in the terminus region does not result from a lack of identifiable coding sequences. We can therefore conclude that the terminus region is composed mainly of expressable, albeit inessential, protein-encoding genes.


Subject(s)
Bacterial Proteins/genetics , DNA Replication , DNA, Bacterial/genetics , Escherichia coli/genetics , Genes, Bacterial , Bacterial Proteins/chemistry , Cloning, Molecular , Molecular Weight , Restriction Mapping
4.
Mol Gen Genet ; 220(2): 341-4, 1990 Jan.
Article in English | MEDLINE | ID: mdl-1691435

ABSTRACT

The sequence of the PcnB protein of Escherichia coli, a protein required for copy number maintenance of ColE1-related plasmids, was compared with the PIR sequence database. Strong local similarities to the sequence of the E. coli protein tRNA nucleotidyltransferase were found. Since a substrate of the latter protein, tRNA, structurally resembles the RNAs that control ColE1 copy number we believe that we may have identified a region in PcnB that interacts with these RNAs. Consistent with this idea is our observation that PcnB is required for the replication of R1, a plasmid whose replication is also regulated by a small RNA.


Subject(s)
Bacterial Proteins/genetics , Bacteriocin Plasmids , Escherichia coli Proteins , Escherichia coli/genetics , Plasmids , Polynucleotide Adenylyltransferase , RNA, Bacterial/metabolism , Amino Acid Sequence , DNA Replication , Molecular Sequence Data , RNA Nucleotidyltransferases/genetics , Sequence Homology, Nucleic Acid
5.
Mol Microbiol ; 3(7): 903-10, 1989 Jul.
Article in English | MEDLINE | ID: mdl-2677604

ABSTRACT

A gene, pcnB, affecting the copy number of ColE1-related plasmids has been cloned and mapped to 3.6 min on the Escherichia coli chromosome between panD and fhu. The gene encodes a previously undescribed 48 kD protein. Several independently isolated mutants exhibiting the same phenotype, reduced copy number, have been shown to be pcnB-.


Subject(s)
Bacterial Proteins/genetics , Escherichia coli/genetics , Genes, Bacterial , Plasmids , Alleles , Ampicillin Resistance , Cloning, Molecular , Gene Expression Regulation , Molecular Weight , Mutation , Phenotype , Protein Biosynthesis , Restriction Mapping , Transcription, Genetic , Transduction, Genetic
6.
Mol Gen Genet ; 214(3): 523-32, 1988 Nov.
Article in English | MEDLINE | ID: mdl-3063949

ABSTRACT

P1 transduces bacterial chromosomal markers with widely differing frequencies. We use quantitative Southern hybridisations here to show that, despite this, most markers are packaged at similar levels. Exceptions are a group of markers near 2 min and another at 90 min which seem to be packaged at levels two- to threefold higher. We thus conclude that certain marker frequency variations in transduction can be explained by differences in packaging level, but that most cannot. The limited range in packaging levels suggests that P1 can initiate the packaging of chromosomal DNA from many sites. This idea is supported by our failure to find any chromosomal sequences with homology to the phage pac site and by the occurrence of hybridising bands which seem to suggest sequential packaging from a large number of specific sites. We eliminate the possibility that chromosomal DNA packaging is the result of endonucleolytic cutting by the P1 res enzyme.


Subject(s)
Bacteriophages/genetics , Capsid/physiology , DNA, Viral/physiology , Escherichia coli/genetics , Transduction, Genetic , Autoradiography , Bacteriophages/physiology , DNA Probes , DNA, Bacterial/analysis , DNA, Viral/analysis , Escherichia coli/analysis , Nucleic Acid Hybridization , Plasmids
7.
Mol Gen Genet ; 202(3): 446-54, 1986 Mar.
Article in English | MEDLINE | ID: mdl-3012270

ABSTRACT

An 8.2 kb fragment of E. coli chromosomal DNA, when cloned in increased copy number, suppresses the dnaA46 mutation, and an abundant protein of about 68 kd (60 kd when measured by us), encoded by the fragment, is essential for the suppression (Takeda and Hirota 1982). Mapping experiments show that the fragment originates from the 94 min region of the chromosome. It encodes several proteins but only one abundant polypeptide of the correct size, the product of the groEL gene. Suppression by the fragment is allele specific; those mutations which map to the centre of the gene are suppressed. Other initiation mutants including dnaA203, dnaA204, dnaA508, dnaAam, dnaC, dnaP and dnaB252 are not suppressed. Most suppressed strains are cold-sensitive suggesting an interaction between the mutant proteins (or their genes) and the suppressing protein or proteins.


Subject(s)
Bacterial Proteins/genetics , DNA Replication , DNA, Bacterial/genetics , Escherichia coli/genetics , Genes, Bacterial , Genes , Heat-Shock Proteins , Mutation , Suppression, Genetic , Chaperonins , Cloning, Molecular , DNA Restriction Enzymes , Escherichia coli Proteins , Genotype , Plasmids , Species Specificity
8.
Planta ; 122(1): 45-52, 1975 Jan.
Article in English | MEDLINE | ID: mdl-24435920

ABSTRACT

Leaf tissue of the panicoid grass Pennisetum purpureum (Schum) and of the chloridoid grass Muhlenbergia montana (Hitchcock) were fractionated to produce preparations enriched in the contents of mesophyll and bundle sheath cells. Sucrose phosphate synthetase and sucrose synthetase were found predominantly in the mesophyll tissues of both species, as was uridine-diphosphate-glucose pyrophosphorylase in Pennisetum. In Muhlenbergia this enzyme was more plentiful in the bundle sheath cell fraction.Starch synthetase and adenosine-diphosphate-glucose pyrophosphorylase were plentiful in bundle sheath extracts of both species but phosphorylase was associated with extracts of mesophyll cells. In no case was there a clear-cut compartmentation of enzymes between leaf fractions.

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