Search results for "Origin of replication"
showing 10 items of 10 documents
Dextransucrase Expression Is Concomitant with that of Replication and Maintenance Functions of the pMN1 Plasmid in Lactobacillus sakei MN1
2017
The exopolysaccharide synthesized by Lactobacillus sakei MN1 is a dextran with antiviral and immunomodulatory properties of potential utility in aquaculture. In this work we have investigated the genetic basis of dextran production by this bacterium. Southern blot hybridization experiments demonstrated the plasmidic location of the dsrLS gene, which encodes the dextransucrase involved in dextran synthesis. DNA sequencing of the 11,126 kbp plasmid (pMN1) revealed that it belongs to a family which replicates by the theta mechanism, whose prototype is pUCL287. The plasmid comprises the origin of replication, repA, repB, and dsrLS genes, as well as seven open reading frames of uncharacterized f…
Replication origins and pause sites in sea urchin mitochondrial DNA
1992
We have used a combination of one- and two-dimensional agarose gel electrophoresis, and solution hybridization to strand-specific probes, to map the replication origin of sea urchin mitochondrial DNA and to investigate the structure of replication intermediates. These assays are consistent with replication initiating unidirectionally from the D-loop region by D-loop expansion, as in vertebrates. A prominent site of initiation of lagging-strand synthesis lies at, or near to, the boundary between the genes for ATPase 6 and COIII, which is also close to a pause site for leading-strand synthesis. These findings suggest a role for pause sites in the regulation of mitochondrial transcription and …
Sequences in the 5′ Nontranslated Region of Hepatitis C Virus Required for RNA Replication
2001
ABSTRACT Sequences in the 5′ and 3′ termini of plus-strand RNA viruses harbor cis -acting elements important for efficient translation and replication. In case of the hepatitis C virus (HCV), a plus-strand RNA virus of the family Flaviviridae , a 341-nucleotide-long nontranslated region (NTR) is located at the 5′ end of the genome. This sequence contains an internal ribosome entry site (IRES) that is located downstream of an about 40-nucleotide-long sequence of unknown function. By using our recently developed HCV replicon system, we mapped and characterized the sequences in the 5′ NTR required for RNA replication. We show that deletions introduced into the 5′ terminal 40 nucleotides abolis…
Molecular Characterization of the Leucine Plasmid from Buchnera aphidicola , Primary Endosymbiont of the Aphid Acyrthosiphon pisum
2000
The complete sequence of the leucine plasmid of Buchnera aphidicola from the aphid Acyrthosiphon pisum (pLeu-BAp) is reported. Its gene organization was concordant with those of other leucine plasmids of Buchnera from aphids of the Aphidini and Macrosiphini tribes. Three inverted repeats are present in pLeu-BAp. Two of them are also present in pLeu from the family Aphididae: (i) SIR1, located downstream the leucine operon, resembles a rho-independent terminator of transcription, and (ii) LIR1, located upstream of the leucine operon, is suggested to be involved in transcription termination or messenger stability. The third, located near the putative ATGC repeats involved in the origin of rep…
A natural A/T-rich sequence from the yeast FBP1 gene exists as a cruciform in Escherichia coli cells.
1993
Abstract Palindromic or semipalindromic sequences can adopt cruciform structures in DNA in vitro. It has been demonstrated in some cases that A/T-rich cruciforms exist also in vivo in Escherichia coli. The biological function of those structures is not understood although putative cruciforms have been found in interesting locations on replication origins, operators, or transcriptional termination regions. Here we show by means of the use of structure-dependent nucleases that the 3′ end of the yeast FBP1 gene contains a stable cruciform both in vitro and in E. coli cells and that in both cases, its extrusion depends on the DNA supercoiling state.
Plasmid conjugation from Proteobacteria as evidence for the origin of xenologous genes in Cyanobacteria
2014
Comparative genomics have shown that 5% of Synechococcus elongatus PCC 7942 genes are of probable proteobacterial origin. To investigate the role of interphylum conjugation in cyanobacterial gene acquisition, we tested the ability of a set of prototype proteobacterial conjugative plasmids (RP4, pKM101, R388, R64, and F) to transfer DNA from Escherichia coli to S. elongatus. A series of BioBrick-compatible, mobilizable shuttle vectors was developed. These vectors were based on the putative origin of replication of the Synechococcus resident plasmid pANL. Not only broad-host-range plasmids, such as RP4 and R388, but also narrower-host-range plasmids, such as pKM101, all encoding MPFT-type IV …
Chromatin structure of transposon Tn903 cloned into a yeast plasmid
1989
Transposon Tn903 contains the APH gene for kanamycin resistance, which is active in yeast [A. Jiménez and J. Davies (1980) Nature (London) 287, 869-871] and is flanked by two inverted repeats (IR) 1057 bp long. When plasmid pAJ50, carrying Tn903 and the 2-microns circle origin of replication, is cloned into Saccharomyces cerevisiae, nucleosomes are assembled in vivo on the prokaryotic DNA of the transposon. Indirect end labeling revealed that three nucleosomes are preferentially positioned on symmetrical sequences from both IRs. DNase I digestion also confirmed that the chromatin structure is symmetrical in both IRs. This suggests that sequence determinants are decisive for chromatin struct…
Biochemistry and Molecular Biology of DNA Replication in Yeast
1985
For the past two decades, the study of the mechanism of DNA replication has been focused mainly on the chromosomes of the simple prokaryotes and their viruses (1). The complexity of the eukaryotic genome and multiple levels of control during the replication of eukaryotic chromosomes have until recently prevented similar studies. In recent years, a lower eukaryote, the yeast Saccharomyces cerevisiae, has become a major focus of efforts in molecular biology. In this chapter, I will briefly review accomplishments in this area. Yeast is an ideal model system for studies on the structure and replication of the eukaryotic chromosome. Yeast cells are easy to grow and study biochemically. Genetic a…
Enhancement of hepatitis C virus RNA replication by cell culture-adaptive mutations.
2001
ABSTRACT Studies of the Hepatitis C virus (HCV) replication cycle have been made possible with the development of subgenomic selectable RNAs that replicate autonomously in cultured cells. In these replicons the region encoding the HCV structural proteins was replaced by the neomycin phosphotransferase gene, allowing the selection of transfected cells that support high-level replication of these RNAs. Subsequent analyses revealed that, within selected cells, HCV RNAs had acquired adaptive mutations that increased the efficiency of colony formation by an unknown mechanism. Using a panel of replicons that differed in their degrees of cell culture adaptation, in this study we show that adaptive…
Viral and cellular determinants of hepatitis C virus RNA replication in cell culture.
2003
Studies on the replication of hepatitis C virus (HCV) have been facilitated by the development of selectable subgenomic replicons replicating in the human hepatoma cell line Huh-7 at a surprisingly high level. Analysis of the replicon population in selected cells revealed the occurrence of cell culture-adaptive mutations that enhance RNA replication substantially. To gain a better understanding of HCV cell culture adaptation, we characterized conserved mutations identified by sequence analysis of 26 independent replicon cell clones for their effect on RNA replication. Mutations enhancing replication were found in nearly every nonstructural (NS) protein, and they could be subdivided into at …