Biomedical subjects

J García-Lara

Publications and source records attributed to J García-Lara.

7 recordsLinked to original sources

PubMed2000

Cell division inhibitors SulA and MinC/MinD block septum formation at different steps in the assembly of the Escherichia coli division machinery.

SulA and MinCD are specific inhibitors of cell division in Escherichia coli. In this paper, size exclusion chromatography was used to study the effect of the SulA and MinCD division inhibitors on the oligomerization state of endogenous FtsZ in cytoplasmic extracts, and immunofluorescence microscopy was used to determine the effect of SulA and MinCD on the formation of FtsZ, FtsA and ZipA rings at potential division sites. SulA prevented the formation of high-molecular-weight FtsZ polymers by interfering with FtsZ dimerization and subsequent oligomerization. In contrast, the MinCD division inhibitor did not prevent the oligomerization of FtsZ in the cell extracts or the formation of FtsZ and ZipA ring structures in vivo. However, MinCD did prevent the formation of FtsA rings. Increased expression of ftsA suppressed MinCD-induced division inhibition, but had no effect on SulA-induced division inhibition. These results indicate that MinCD blocks the assembly of the septation machinery at a later step than SulA, at the stage at which FtsA is added to the FtsZ ring.

Adenosine Triphosphatases↗

PubMed2000

The role of genomics in approaching the study of Borrelia DNA replication.

The identification of chromosomal and episomal origins of replication in the genome of the causative agent of Lyme disease, the spirochete Borrelia burgdorferi, has been greatly facilitated by genomics. Analysis of genome features, including strand compositional asymmetries, organizational similarities to other bacterial origins of replication, and the presence of homologues of genes involved in replication and partitioning, have contributed to the identification of a collection of putative origins of replication within the Borrelia genome. This analysis has provided the basis for the experimental verification of origins in the linear chromosome and in the linear plasmid Ip28-2. Information generated during the study of these origins will significantly contribute to the understanding of the mechanisms of replication and partitioning in Borrelia.

Amino Acid Sequence↗

PubMed1999

Bacterial cell division.

Formation of the bacterial division septum is catalyzed by a number of essential proteins that assemble into a ring structure at the future division site. Assembly of proteins into the cytokinetic ring appears to occur in a hierarchial order that is initiated by the FtsZ protein, a structural and functional analog of eukaryotic tubulins. Placement of the division site at its correct location in Escherichia coli requires a division inhibitor (MinC), that is responsible for preventing septation at unwanted sites near the cell poles, and a topological specificity protein (MinE), that forms a ring at midcell and protects the midcell site from the division inhibitor. However, the mechanism responsible for identifying the position of the midcell site or the polar sites used for spore septum formation is still unclear. Regulation of the division process and its coordination with other cell cycle events, such as chromosome replication, are poorly understood. However, a protein has been identified in Caulobacter (CtrA) that regulates both the initiation of chromosome regulation and the transcription of ftsZ, and that may play an important role in the coordination process.

Cell Division↗

PubMed1997

Exogenous isolation of Hgr plasmids from coastal Mediterranean waters and their effect on growth and survival of Escherichia coli in sea water.

Four self-conjugative plasmids conferring mercury resistance were exogenously isolated from coastal Mediterranean sea water. All the plasmids isolated (93.2, 73.1, 65.2 and 221 kb) showed narrow-spectrum mercury resistance, and no UV resistance; three of them conferred cadmium resistance (Cdr 1.5 mM) and two of them encoded streptomycin resistance (50 micrograms/ml). Their influence on the survival of the recipient strain Escherichia coli JGL1 in different seawater media was studied and compared with that of two plasmids previously isolated by the same procedure from an epilithic community of the Bristol Channel, Great Britain. A significant negative influence was observed for the largest (221 kb) of the plasmids isolated and one of the epilithic plasmids (239 kb). The effect of the plasmids on E.coli JGL1 was dependent on the seawater media.

Cadmium↗

PubMed1996

An extracellular factor regulates expression of sdiA, a transcriptional activator of cell division genes in Escherichia coli.

The sdiA gene codes for a protein that regulates expression of the ftsQAZ cluster of essential cell division genes of Escherichia coli. SdiA up-regulates the ftsQ2p promoter that initiates transcription into the ftsQAZ cluster. In this paper, we report that expression of sdiA is itself regulated by a factor that is released into the growth medium by E. coli. When medium that had previously supported growth of E. coli (conditioned medium) was used to support growth of an indicator E. coli strain that contained an sdiA-lacZ transcriptional reporter, there was a 50 to 80% decrease in sdiA expression as monitored by beta-galactosidase activity. The down-regulation of PsdiA was associated with a decrease in expression of the SdiA target promoter ftsQ2p, as monitored by expression of an ftsQ2p-lacZ transcriptional fusion. An effect of conditioned medium on ftsQ2p expression was not seen when the wild-type sdiA gene was disrupted by insertional mutagenesis, indicating that the effect on ftsQ2p expression was secondary to the down-regulation of PsdiA. Conditioned medium had no effect on expression of Plac, PrpoS, or several other promoters associated with the ftsQAZ gene cluster (ftsQ1p and ftsZ1-4p). This suggests that the response is specific for PsdiA and for promoters that are regulated by the sdiA gene product and that cell-to-cell signalling may play a role in regulating expression of this group of genes.

4-Butyrolactone↗

PubMed1993

Effect of previous growth conditions on the starvation-survival of Escherichia coli in seawater.

The starvation-survival of Escherichia coli in seawater was assessed by plate and epifluorescence counts, 3H-label decrease, cellular DNA concentrations, and metabolic activities. These assays were performed on two types of populations, adapted and non-adapted to seawater. The number of viable cells in the adapted population remained constant throughout starvation-survival in sterile seawater. In contrast, a significant decrease in the ability of the non-adapted E. coli to form colonies on plates following starvation-survival in sterile seawater was observed. However, this drop in viable counts was not mirrored by the epifluorescence counts and 3H-label, which did not show major changes for either population during the experiments, indicating maintenance of the number of cells. In addition, a significant increase in and subsequent maintenance of DNA content and thymidine incorporation was observed for both populations during starvation-survival in sterile seawater. The changes in cell-attached exoproteolytic activity and electron transport system activity showed that adapted and non-adapted E. coli cells maintain their metabolic potential. Cell-free exoproteolytic activity was drastically reduced in both populations. Adapted cells showed higher electron transport system activity and thymidine incorporation than non-adapted cells at the onset of starvation-survival. The effect of previous adaptation on E. coli starvation-survival, as assessed by plate counts and 3H-label decrease, was also observed in raw seawater. It seems from these data that the biological potential of E. coli cells suspended in sterile seawater has not been switched off or impaired seriously.

Acridine Orange↗

PubMed1991

Mortality of marine bacterial strains in seawater.

As an approach for assessing the dynamics of bacterial population in seawater, the survival of five isolated marine bacteria strains was assessed by the disappearance of radioactivity in the cold trichloroacetic acid (TCA)-insoluble fraction from a previously 3H-labeled culture. Metabolic activity during survival experiments was assessed by the measurement of electron transport system (ETS) activity. Fractionated filtration was used to assess the grazing mortality. The particulate fraction that passed 2.0 microns and was retained in 0.2 microns was the main cause of mortality.

Bacteria↗