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J D Keasling

Publications and source records attributed to J D Keasling.

8 recordsLinked to original sources
PubMed1994

Polyphosphate metabolism in Escherichia coli.

Polyphosphate metabolism in Escherichia coli was studied in order to determine the role of polyphosphates in energy and phosphate metabolism. Phosphate-shift experiments were performed on wild-type E. coli W3110 and on an E. coli strain mutant in the genes encoding the polyphosphate-metabolizing enzymes polyphosphate kinase (PPK) and polyphosphatase (PPX). The levels of polyphosphates were measured by [31P]NMR, and the activities of PPK and PPX were measured using enzymatic assays. During phosphate starvation, the intracellular level of polyphosphate was not detectable in E. coli W3110; the activities of PPX and alkaline phosphatase were high relative to those during exponential growth. During the shift from phosphate starvation to phosphate surplus conditions, PPX activity decreased and PPK activity and intracellular polyphosphate stores increased dramatically. These results imply an important role for polyphosphates in cellular energy and phosphate storage and in adaptation to adverse growth conditions.

Acid Anhydride Hydrolases↗
PubMed1992

Replication of prophage P1 is cell-cycle specific.

P1 prophage replication during the Escherichia coli division cycle has been analyzed by using the membrane-elution technique to produce cells labelled at different times during the division cycle and scintillation counting for quantitative analysis of radioactive prophage DNA. P1 prophage replicates during a restricted portion of the bacterial division cycle, like the minichromosome, but at a time during the division cycle different than the time at which the minichromosome replicates in the same cell. A high-copy mini-R6K plasmid present in the same cell replicates throughout the division cycle. Over a wide range of growth rates, the P1 prophage replicates approximately one-half generation after the minichromosome replicates. Thus, the mechanisms underlying P1 replication are similar to those for the F plasmid and the chromosome. Replication occurs when some property related to cell size or cell mass reaches a constant value per origin.

Cell Cycle↗
PubMed1992

Replication of the R6K plasmid during the Escherichia coli cell cycle.

The cell-cycle replication pattern of the R6K plasmid has been investigated by using the membrane-elution technique to produce cells labelled at different times during the division cycle and scintillation counting for quantitative analysis of radioactive plasmid DNA. The high-copy plasmid R6K replicates exponentially in a cell-cycle-independent manner. A mini-R6K plasmid deleted for the ori alpha origin of replication also replicates, exponentially in a cell-cycle-independent manner.

Cell Cycle↗
PubMed1991

Cell-cycle-specific F plasmid replication: regulation by cell size control of initiation.

F plasmid replication during the Escherichia coli division cycle was investigated by using the membrane-elution technique to produce cells labeled at different times during the division cycle and scintillation counting for quantitative analysis of radioactive plasmid DNA. The F plasmid replicated, like the minichromosome, during a restricted portion of the bacterial division cycle; i.e., F plasmid replication is cell-cycle specific. The F plasmid replicated at a different time during the division cycle than a minichromosome present in the same cell. F plasmid replication coincided with doubling in the rate of enzyme synthesis from a plasmid-encoded gene. When the cell cycle age of replication of the F plasmid was determined over a range of growth rates, the cell size at which the F plasmid replicated followed the same rules as did replication of the bacterial chromosome--initiation occurred when a constant mass per origin was achieved--except that the initiation mass per origin for the F plasmid was different from that for the chromosome origin. In contrast, the high-copy mini-R6K plasmid replicated throughout the division cycle.

Cell Cycle↗
PubMed1990

The regulatory mechanisms of human immunodeficiency virus replication predict multiple expression rates.

The qualitative nature of human immunodeficiency virus replication dynamics is examined by using the known action of regulatory proteins. The combination of activation of transcription by the Tat protein and the influence of the Rev protein on processing of RNA leads to a regulatory loop that can have multiple expression rates. In the lower state, the regulatory loop leads to low, or no, production of virus, which corresponds to its quiescent state. Conversely, when the regulatory loop is in the upper state, active production of virus is maintained over time. These features of the kinetic model have implications for both human immunodeficiency virus-related illness and therapeutics and predict testable experimental results in vitro.

Gene Products, tat↗
PubMed1989

ColE1 plasmid replication: a simple kinetic description from a structured model.

Time constant analysis illustrates how a complex model for ColE-1-type plasmid replication can be reduced to a simple, realistic description of the overall replication kinetics. The analysis leads to the identification of the key parameters governing plasmid replication and to the estimation of parameters that are difficult to determine experimentally. The overall kinetics of ColE1-type plasmid replication are zerothorder in production and first order in dilution. Experimental data confirm our predictions. Similar analysis of the postulated mechanism for the action of the Rom protein reveals that it has no significant effect on plasmid replication dynamics.

Cell Cycle↗
PubMedDate not supplied

Replication of mini-F plasmids during the bacterial division cycle.

The cell-cycle replication patterns of two mini-F plasmids have been examined using the membrane-elution technique (to produce cells labelled at different times during the division cycle) and scintillation counting (for quantitative analysis of radioactivity incorporated into plasmid DNA). The mini-F plasmid pML31, which contains the oriV and oriS origins of replication, replicates in a cell-cycle-specific manner with a pattern and cell-cycle timing similar to the parental F plasmid. The mini-F plasmid pMF21, deleted for the region containing the oriV origin of replication, replicates more randomly throughout the division cycle. These results suggest that the oriV origin of replication may be related to cell-cycle-specific replication of the F plasmid.

Cell Division↗