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R A Bender

Publications and source records attributed to R A Bender.

At least 55 records · Page 3Linked to original sources
PubMed1988

Bidirectional promoter in the hut(P) region of the histidine utilization (hut) operons from Klebsiella aerogenes.

The hut(P) region (i.e., the region responsible for regulation of hutUH expression) of the Klebsiella aerogenes histidine utilization (hut) operons contains a bidirectional promoter. One transcript from this promoter encodes the hutUH operon; the role of the oppositely directed transcript is unknown, although it appears to be involved in regulating hutUH expression (A.J. Nieuwkoop, S.A. Boylan, and R.A. Bender, J. Bacteriol. 159:934-939, 1984). A 247-base-pair (bp) fragment containing hut(P) carries two RNA-polymerase-binding sites agree with the start sites of the two transcripts produced from hut(P) DNA in vitro and in vivo. The binding sites share a 4-bp region, suggesting that occupancy of the regulatory site precludes occupancy of the hutUH promoter, and vice versa. In the absence of positive effectors, the binding to the site responsible for hutUH transcription is weaker than the binding to the site responsible for regulation. The nucleotide sequence of the 250-bp fragment containing hut(P) contains two possible matches to the consensus sequence for Escherichia coli promoters, a better and worse match, corresponding in position to the stronger and weaker RNA-polymerase-binding sites, respectively. The sequence also contains a region similar to the consensus sequence for binding of the catabolite gene activator protein of E. coli. A sequence similar to the consensus for Ntr-dependent promoters was also found, overlapping both RNA-polymerase-binding sites, but it is not a functional promoter. Finally, an initiation codon preceded by a Shine-Dalgarno consensus sequence and followed by an open reading frame identifies a probable start of the hutU gene coding sequences.

Base Sequence↗
PubMed1988

Klebsiella pneumoniae origin of replication (oriC) is not active in Caulobacter crescentus, Pseudomonas putida, and Rhodobacter sphaeroides.

A DNA fragment carrying genes encoding the conjugal transfer system of the broad host range plasmid RK2 was inserted into a plasmid carrying the chromosomal origin of replication (oriC) from Klebsiella pneumoniae. The resulting plasmid, pEON1, was readily transferred between gram-negative bacteria and carried two potential origins of replication: oriC and the replication origin from pBR322 (oriPBR). Although pEON1 could be transferred to Caulobacter crescentus, Pseudomonas putida, and Rhodobacter sphaeroides, pEON1 was not maintained in these strains. However, an oriC-containing plasmid was maintained in these nonenteric bacteria when an RK2 origin of replication was present on the plasmid. Thus, the inability of pEON1 to be established in a nonenteric bacterium represents a failure of oriC to function as an origin of replication rather than a toxic effect of oriC. The initiation potential of the chromosomal origin of replication from K. pneumoniae appears to be realized only in enteric bacteria.

Bacteria↗
PubMed1988

Vinca alkaloids.

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Animals↗
PubMed1987

Periodic synthesis of phospholipids during the Caulobacter crescentus cell cycle.

Net phospholipid synthesis is discontinuous during the Caulobacter crescentus cell cycle with synthesis restricted to two discrete periods. The first period of net phospholipid synthesis begins in the swarmer cell shortly after cell division and ends at about the time when DNA replication initiates. The second period of phospholipid synthesis begins at a time when DNA replication is about two-thirds complete and ends at about the same time that DNA replication terminates. Thus, considerable DNA replication, growth, and differentiation (stalk growth) occur in the absence of net phospholipid synthesis. In fact, when net phospholipid synthesis was inhibited by the antibiotic cerulenin through the entire cell cycle, both the initiation and the elongation phases of DNA synthesis occurred normally. An analysis of the kinetics of incorporation of radioactive phosphate into macromolecules showed that the periodicity of phospholipid synthesis could not have been detected by pulse-labeling techniques, and only an analysis of cells prelabeled to equilibrium allowed detection of the periodicity. Equilibrium-labeled cells also allowed determination of the absolute amount of phosphorus-containing macromolecules in newborn swarmer cells. These cells contain about as much DNA as one Escherichia coli chromosome and about four times as much RNA as DNA. The amount of phosphorus in phospholipids is about one-seventh of that in DNA, or about 3% of the total macromolecular phosphorus.

Cell Cycle↗
PubMed1987

Vinca alkaloids.

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Animals↗
PubMed1985

Recombination deficient mutant of Caulobacter crescentus.

A recombination-deficient (Rec-) strain of Caulobacter crescentus has been isolated from a collection of mutants sensitive to ultraviolet irradiation. The Rec- mutant fails to give recombinants following phi Cr30-mediated generalized transduction or following RP4-mediated conjugation. The recombination frequency in the Rec- strain is at least 5000-fold lower than in the wild type strains. The Rec- mutant is indistinguishable from wild type in terms of morphology, growth rate, viability, and phage sensitivities, differing only in properties known to be associated with recA-type mutations in other organisms: recombination frequency, ultraviolet sensitivity, and Weigle reactivation. The map location of the rec-526 allele has not been identified, but rec-526 can be cotransferred with the fla-169 mutation by RP4-mediated conjugation at low frequency. This apparent linkage has been used to move the rec mutation to other strains. The Rec- mutant resembles recA strains of other organisms and provides a healthy strain severely deficient in recombination for use in complementation and cloning studies involving C. crescentus.

Conjugation, Genetic↗
PubMed1984

A restriction enzyme cleavage map of the histidine utilization (hut) genes of Klebsiella aerogenes and deletions lacking regions of hut DNA.

The histidine utilization (hut) operons of Klebsiella aerogenes were cloned into pBR322. The hut genes are wholly contained on a 7.9 kilobase pair fragment bounded by HindIII restriction sites and expression of hut is independent of the orientation of the fragment with respect to pBR322. A restriction map locating the 27 cleavage sites within hut for the enzymes, HindIII, PvuII, SalI, BglII, KpnI, PstI, SmaI, AvaI, and BamHI was deduced. Several of the cleavage sites for the enzymes HaeIII and HinfI were also mapped. A set of deletion plasmids was isolated by removing various restriction fragments from the original plasmid. These deletions were characterized and were used to assist in mapping restriction sites. This physical characterization of hut DNA opens the way for genetic and molecular analysis of the regulation of hut gene expression in vitro as well as in vivo.

Base Composition↗
PubMed1984

Genetic and physical maps of Klebsiella aerogenes genes for histidine utilization (hut).

Deletion derivatives of the hut-containing plasmid pCB101 were tested against point mutants defective in individual genes of the histidine utilization (hut) operons using a complementation/recombination assay. Location of the genes of the right operon, hutU and hutH, was confirmed by direct assay of the gene products, urocanase and histidase; location of the repressor gene was identified by measuring the ability of the plasmid-carried genes to repress the formation of histidase from a chromosomal location. The analysis of eight deletion plasmids unambiguously confirms the map order of the hut genes as hutI-G-C-U-H, and demonstrates that, in Klebsiella aerogenes, the hutU and hutH genes are transcribed from their own promoter. In addition, the genetic map of hut can be aligned with the restriction map of the hut DNA in plasmid pCB101. One of the deletion plasmids studied apparently encodes a defective histidase subunit that is trans-dominant to active histidase. Another deletion, which completely removes the left operon, hutIG, allows high level expression of the hutUH operon and thus overproduction of a toxic intermediate.

Escherichia coli↗
PubMed1984

Ultraviolet mutagenesis and inducible DNA repair in Caulobacter crescentus.

The ability to reactivate ultraviolet (UV) damaged phage phi CbK (W-reactivation) is induced by UV irradiation of Caulobacter crescentus cells. Induction of W-reactivation potential is specific for phage phi CbK, requires protein synthesis, and is greatly reduced in the presence of the rec-526 mutation. The induction signal generated by UV irradiation is transient, lasting about 1 1/2-2 h at 30 degrees C; if chloramphenicol is present during early times after UV irradiation, induction of W-reactivation does not occur. Induction is maximal when cells are exposed to 5-10 J/m2 of UV, a dose that also results in considerable mutagenesis of the cells. Taken together, these observations demonstrate the existence of a UV inducible, protein synthesis requiring, transiently signalled, rec-requiring DNA repair system analogous to W-reactivation in Escherichia coli. In addition, C. crescentus also has an efficient photoreactivation system that reverses UV damage in the presence of strong visible light.

Bacteria↗
PubMed1984

Use of bacteriophage P1 as a vector for Tn5 insertion mutagenesis.

Infection of a strain lysogenic for bacteriophage P1 CM with P1::Tn5 followed by simultaneous selection for the chloroamphenicol resistance associated with the resident prophage and the kanamycin resistance associated with Tn5 results in a large number of independent Tn5 insertion mutations. This superinfection-selection protocol is a fast, easy, and safe way to isolate null mutations in enteric bacteria without generating unwanted cryptic mutations elsewhere in the genome.

Bacteriophages↗
PubMed1984

Transposon Tn5 encodes streptomycin resistance in nonenteric bacteria.

Strains of Caulobacter crescentus, Pseudomonas putida, Acinetobacter calcoaceticus, Rhizobium meliloti, and Rhodopseudomonas sphaeroides carrying the kanamycin resistance-encoding transposon Tn5 were 15 to 500 times more resistant to streptomycin than transposon-free strains. The streptomycin resistance determinant, which is separable from the kanamycin resistance determinant of Tn5, was not expressed in Escherichia coli or Klebsiella aerogenes.

Bacteria↗
PubMed1984

Regulation of hutUH operon expression by the catabolite gene activator protein-cyclic AMP complex in Klebsiella aerogenes.

RNA polymerase transcribed the hutUH operon of Klebsiella aerogenes if the catabolite gene activator protein (CAP) and cyclic AMP (cAMP) were present or if the DNA template was derived from a promoter mutant in which hutUH expression was independent of the need for positive effectors. In the absence of CAP or cAMP, not only was hutUH transcription absent, but transcription in the opposite direction (toward hutC) was initiated at a site (pC) ca. 70 base pairs from the site (pUH) of hutUH mRNA initiation. When the pC promoter was cloned in front of a promoterless galK gene, active expression of galK was observed. Thus, the pC promoter is active in vivo as well as in vitro. Transcription from pUH and pC may be mutually exclusive, with the major effect of CAP and cAMP being to prevent transcription from pC, thus relieving the antagonistic effect on transcription from pUH. This "double-negative" control by CAP-cAMP is supported by two observations: (i) CAP-cAMP was unable to activate transcription from pUH if RNA polymerase had been previously bound to pC and (ii) a mutation that allowed transcription from pUH in the absence of positive effectors simultaneously eliminated the activity of pC. An alternative model, in which CAP-cAMP is required for pUH expression and RNA polymerase binding at pC serves to modulate this control in some unknown way, is also considered. The physiological role of the transcript from pC other than regulation of pUH is unknown.

Base Sequence↗
PubMed1983

Recombination-induced suppression of cell division following P1-mediated generalized transduction in Klebsiella aerogenes.

Klebsiella aerogenes recombinants resulting from bacteriophage P1-mediated generalized transduction failed to increase in number for approximately six generations after transduction. Nevertheless these recombinants continued to grow and became sensitive to penicillin after a transient resistance, suggesting that the cells were growing as long, non-dividing filaments. When filamentous cells were isolated from transduced cultures by gradient centrifugation, recombinants were 1000-fold more frequent among the filaments than among the normal-sized cells. The suppression of cell-division lasted for six generations whether markers near the origin (gln, ilv) or terminus (his, trp) of chromosome replication were used, despite a 50-fold difference in transduction frequencies for these markers. The suppression of cell division was a host response to recombination rather than to P1 invasion since cells lysogenized by P1 in these same experiments showed only a short (two generation) suppression of cell division. We speculate that the suppression of cell-division is an SOS response triggered by the degraded DNA not incorporated in the final recombinant. We demonstrate that both the filamentation and the transient penicillin resistance of recombinant cells can be exploited to enrich greatly for recombinants, raising transduction frequencies to as high as 10(-3).

Bacteriophages↗
PubMed1983

Nitrogen regulation system of Klebsiella aerogenes: the nac gene.

In Klebsiella aerogenes, the product of a his-linked gene, nac, appears to play a crucial role in tying the synthesis of enzymes activated or repressed by ammonia deprivation, such as histidase and glutamate dehydrogenase, to the known regulators of nitrogen assimilation, the products of glnG and glnF.

Ammonia↗
PubMed1982

A nonsense mutation in the structural gene for glutamine synthetase leading to loss of nitrogen regulation in Klebsiella aerogenes.

An amber mutation (glnA3711), the first nonsense mutation isolated in Klebsiella aerogenes, is described. When amber suppressors were present, the mutant made active glutamine synthetase which was more thermolabile than wild type, showing that glnA3711 lies in the structural gene for glutamine synthetase. Strains carrying the glnA3711 allele were unable to express nitrogen regulation of genes coding for histidase, asparaginase, and glutamate dehydrogenase unless amber suppressors were also present. These results support a model that expression of gene(s) from the glnA promoter is required for nitrogen regulation in K. aerogenes.

Genes↗
PubMed1981

Improved generalized transducing bacteriophage for Caulobacter crescentus.

Caulobacter phage phi Cr30T, a temperature-sensitive derivative of the lytic, generalized transducing phage phi Cr30, was isolated as a double temperature-sensitive recombinant in a cross between phi Cr30ts1 and phi Cr30ts2, phi Cr30T mediated generalized transduction of Caulobacter crescentus at frequencies comparable to those of phi Cr30 and eliminated the requirement for irradiation of transducing lysates to prevent killing of transductants on the plate.

Bacteria↗
PubMed1980

Interaction of albumin and vincristine with a human lymphoblastic leukemia cell line in vitro.

To investigate the potential influence of albumin on the antitumor effect of vincristine (VCR), cells from a human lymphoblastic leukemia cell line (CEM) were exposed in vitro to albumin and a clinically attainable VCR concentration (10(-7)M). Increasing concentrations of albumin between 1 and 4 g/100 ml resulted in a corresponding reduction of VCR-induced growth suppression. Studies employing tritiated vincristine ([3H]-VCR) revealed a progressive inhibition of uptake of the radio-label into CEM cells with increasing concentrations of albumin. These in vitro data show a previously unrecognized protective effect of increasing concentrations of albumin on the growth of human lymphoblastic leukemia cells exposed to VCR as a consequence of inhibition of drug uptake. Further in vivo pharmacologic studies will be necessary to determine their clinical significance.

Biological Transport↗