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Biomedical subjects

M Achtman

Publications and source records attributed to M Achtman.

At least 109 records · Page 6Linked to original sources
PubMed1979

Two Escherichia coli chromosomal cistrons, sfrA and sfrB, which are needed for expression of F factor tra functions.

Twelve mutants of Escherichia coli K-12 have been isolated which carry chromosomal mutations that exhibit pleiotropic effects on the expression of F factor tra cistrons. F pilus synthesis, deoxyribonucleic acid transfer, and surface exclusion are all inhibited. Six of the mutants carry sfrA mutations, and six carry sfrB mutations. sfrA and sfrB are cistrons mapping near thr and metE, respectively. Several F-like plasmids are dependent on sfrA and on sfrB for expression of tra cistrons. Plasmids of incompatibility groups C and S are only dependent on sfrB,and other conjugative plasmids are dependent on neither. sfrB mutations also result in changes in certain cell envelope properties, including change sensitivity to certain bacteriophages which use lipopolysaccharide as a receptor, synthesis of nonfunctional flagella, and altered sensitivity to antibiotics.

Chromosome Mapping↗
PubMed1978

The control region of the F sex factor DNA transfer cistrons: restriction mapping and DNA cloning.

A restriction endonuclease map of EcoRI fragment f6 of F sex factor DNA was constructed and aligned with pre-existing physical and genetic maps. Results of genetic complementation tests and analysis of proteins synthesized in minicells from PstI and BglII sub-fragment clones, or from a specific BglII fragment deletion, have allowed mapping of the locations of the origin of DNA transfer and many of the transfer genes known to lie on f6. The proteins detected account for 78% of the coding capacity of fragment f6.

Chromosome Mapping↗
PubMed1978

Fertility repression of F-like conjugative plasmids: physical mapping of the R6--5 finO and finP cistrons and identification of the finO protein.

The locations of the fertility inhibition genes finO and finP of the F-like conjugative multiple antibiotic-resistance plasmid R6-5 have been determined. As found previously for that of the fertility plasmid F, the finP gene of R6-5 is located close to the origin of DNA transfer, oriT, and to the promoter-proximal segment of the tra operon. Thus, finP is close to the site of action of the FinOP fertility inhibition system. In contrast, the finO gene is located on the other side of the tra operon, greater than 35 kilobases from the finP gene; finO is very close to the origin of vegetative replication, oriV, and to cistrons encoding functions involved in autonomous plasmid replication and plasmid incompatibility. A 4.5-kilobase fragment of R6-5 DNA containing the finO gene has been cloned on the high-copy amplifiable vector plasmid pBR322. This hybrid plasmid, designated pKTO31, causes severe repression of conjugal transfer of plasmid F, indicating the production of high cellular levels of finO protein. Two independent finO mutant derivatives were obtained after mutagenesis of the pKTO31 plasmid. Comparison of proteins synthesized by minicells carrying finO(-) mutant plasmids with those carrying various finO(+) plasmids enables the finO gene product to be tentatively identified as a 22,000-dalton protein.

Bacterial Proteins↗
PubMed1978

Cell-cell interactions in conjugating Escherichia coli: purification of F pili with biological activity.

A mutant of the F sex factor has been isolated that produces more F pili per cell than does the wild-type F factor. F pili have been purified in milligram amounts from cells carrying either the mutant or the wild-type sex factor. The technique described yields F pili of up to 99% purity that can specifically bind to Escherichia coli cells and that bind to and reversibly inactivate male-specific bacteriophages. The F pilin subunit has a molecular weight of 10,750 and purified F pili have a buoyant density of 1,200 g/cm3.

Bacterial Proteins↗
PubMed1978

Assignment of tra cistrons to EcoRI fragments of F sex factor DNA.

We describe here the cloning of single EcoRI fragments from the tra region of F DNA using ColE1::Tn3 as vector. These plasmids, as well as the series of Skurray et al. (Proc. Natl. Acad. Sci. U.S.A. 73:64-68, 1976), have been used to refine the map positions of tra cistrons on the F factor as well as to define a new DNA transfer cistron, traM. The current map of the tra cistrons is presented. None of the known tra cistrons, with the exception of traG, straddles an EcoRI site. The EcoRI site at 82 kilobases splits the traG cistron into two portions, an operator-proximal portion necessary for F pilus synthesis and an operator distal portion involved in conjugation itself. The operon structure of the tra cistrons was reevaluated, and we found that traI is at least partially independent of transcription of the traA to traD operon.

Conjugation, Genetic↗
PubMed1978

Cell-cell interactions in conjugating Escherichia coli: role of F pili and fate of mating aggregates.

Bacterial conjugation between Escherichia coli cells was investigated by a combination of physical and genetic techniques, using Hfr, F', or R+ donors and F- recipients. DNA transfer occurred in mating aggregates of up to 50 cells. Multiple interactions between donor and recipient cells occurred, and both F- pilus connections and wall-to-wall contacts were detectable. The detectable F- pilus contacts could be destroyed without either disrupting the mating aggregates or preventing DNA transfer. Hfr X F- mating aggregates did not disaggregate even though recombinant frequencies were inversely proportional to the distance from the origin of DNA transfer. F' or R+ donors formed mating aggregates with F- cells which disaggregated soon after transfer of the autonomous sex factor DNA.

Cell Communication↗
PubMed1977

Conjugation proteins encoded by the F sex factor.

Chimaeric plasmids carrying EcoRI fragments of the F sex factor have been used to identify proteins involved in conjugation and to assign them to tra cistrons. Most of these proteins are incorporated into the cell envelope and are individually regulated at the post-transcriptional level.

Bacterial Proteins↗
PubMed1977

Cell--cell interactions in conjugating Escherichia coli: role of traT protein in surface exclusion.

Escherichia coli cells carrying the F sex factor are poor recipients in conjugation. This phenomenon is called surface exclusion. Two F cistrons, traS and traT, are independently responsible for part of the whole mechanism. The traS gene product reduces DNA transfer within stable mating aggregates. The traT gene product, pTraT, results in a greatly reduced ability to form stable mating aggregates, and thus also leads to reduced DNA transfer within the cell population. pTraT is a 25,000-dalton protein incorporated into the cell envelope outer membrane. It is found in 29,000-84,000 copies per cell, depending on the plasmid expressing it. There is a parallel variation in recipient ability. Models for surface exclusion are discussed.

Bacterial Proteins↗
PubMed1975

Mating aggregates in Escherichia coli conjugation.

Mating mixtures of Escherichia coli cells were shown to contain mating aggregates of two to 20 cells each rather than only mating pairs of two cells each. The mating aggregate size distribution shows two broad peaks, at two to four cells and at eight to 13 cells. The quantitative mating aggregate size distribution and the proportion of male cells in mating aggregates are dependent on the input ratio of male to female cells. At an input ratio of one to one, the average mating aggregate contains equal proportions of male and female cells and most of the cells involved in mating are in large aggregates of seven or more cells each. The deoxyribonucleic acid (DNA) transfer efficiency per mating aggregate cell was constant regardless of average aggregate size or of the ratio of male to female cells in the aggregate. Under optimal conditions essentially every male cell or every female cell in a mating aggregate can be involved in DNA transfer. A comparison of light microscopy, sucrose gradient centrifugation, and analysis with a modified Coulter counter indicated that the number of cells in mating aggregates is best equantitated using a modified Coulter counter.

Cell Aggregation↗
PubMed1972

Conjugational complementation analysis of transfer-deficient mutants of Flac in Escherichia coli.

A series of 102 transfer-deficient (tra(-)) mutants of Flac (84 of which have been previously described) were classified as carrying frameshift, amber, ochre, UGA, and nonsuppressible mutations. Techniques were evolved for using cultures of F prime strains as efficient recipients in matings, for measuring the number of (F'/F') heterozygote cells in transient populations, and for measuring complementation between different Flac tra(-) mutants. These techniques were used to define nine tra cistrons and to assign most of the tra(-) mutations to one or other of these.

Bacteriological Techniques↗
PubMed1972

Genetic analysis of transfer by the Escherichia coli sex factor F, using P1 transductional complementation.

P1 transduction has been used to perform a complementation analysis of a series of transfer-deficient mutants of Flac. The results define ten cistrons and are consistent with the results of a conjugational analysis presented in an accompanying report. Both sets of results are summarized here. Between them, they define eleven cistrons, traA through traK, necessary for conjugational deoxyribonucleic acid (DNA) transfer. Mutants in traI and traD and some in traG still make F-pili, although traD mutants are resistant to f2 phage; their products may be involved in conjugational DNA metabolism. Other mutants in traG and all mutants in the remaining eight cistrons do not make F-pili. One of these, traJ, may be a control cistron, and the others may specify a biosynthetic pathway responsible for synthesis and modification of the F-pilin subunit protein and its assembly into the F-pilus.

Coliphages↗
PubMed1972

Deletion map of the Escherichia coli K-12 sex factor F: the order of eleven transfer cistrons.

A series of Hfr deletion mutants was isolated. These mutants contain deletions which extend from a lambda prophage into an Flac which is integrated into the gal operon. Transfer-deficient deletion mutants were found to fall into four different phenotypic groups when tested for male- and female-specific phage resistance. Conjugational and transductional complementation tests with Flac point mutants deficient in transfer (tra(-)) were performed, and the order of 11 tra cistrons was determined. The tra genes are all located between an F gene for the inhibition of female-specific phages and the transposed lac operon originally carried by the Flac. The order of genes in the Hfr studied was established to be: proC... phi(II) (R)... traJ traA traE traK traB traC traF traH traG traD traI...lac...attlambda...bio.

Chromosome Mapping↗
PubMed1971

Beginning a genetic analysis of conjugational transfer determined by the F factor in Escherichia coli by isolation and characterization of transfer-deficient mutants.

Eighty-four transfer-deficient mutants of Flac have been isolated; 27 of these bear amber mutations and 1 mutant is temperature-sensitive. All the mutants transfer between 10(-2) and <10(-5)% as well as wild-type Flac, all are curable by acridine orange treatment, and all are resistant to the female-specific phage phi(II). Some of the mutants are partially sensitive to female-specific phage tau. Sixty-three of the mutants are resistant to the male-specific phages f1, f2, and Qbeta; 15 are resistant only to f2; and 6 are sensitive to all three male-specific phages. Most of the mutants are still poor recipients in conjugation, but four of the mutants resistant to f1, f2, and Qbeta have become good recipients. Those mutants resistant to all three male-specific phages do not seem to make F-pili.

Acridines↗