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1.
Retraction of F Pili   总被引:24,自引:15,他引:9       下载免费PDF全文
The disappearance of F pili on Escherichia coli cells in the presence of 10−2 M NaCN was studied by electron microscopy and serum-blocking power. The pili which disappeared from the cell did not appear as free pili in the culture medium, suggesting that the pili had retracted into the cell. New pili were produced at a normal rate approximately 3 min after NaCN was removed. The adsorption of either F pili antibody or R17 bacteriophage to the sides of pili and temperatures below 24 C prevented retraction. The disappearance of pili was accompanied by a loss in the ability of cells to adsorb R17 phage and the type of F pili antibody that inhibits R17 phage infection and mating. The ability to adsorb M13 phage and the type of F pili antibody that inhibits M13 phage infection was not affected. This suggests that the tips of retracted pili are exposed.  相似文献   

2.
The extent of removal at various blending speeds (blending spectrum) and the kinetics of reappearance after blending of the ability of male Escherichia coli bacteria to form mating pairs, to adsorb and be infected by ribonucleic acid male phage, and to adsorb and be infected by deoxyribonucleic male phage were identical to the blending spectrum and reappearance kinetics of microscopically visible F pili. The same results were obtained with an Hfr (high-frequency recombinant), F', or resistance transfer factor (RTF) fi(+) mating system. Blending did not affect the viability, growth rate, ability to adsorb T4 phage, or ability to produce new F pili at any of the speeds used. It can be concluded that microscopically visible F pili are an absolute requirement for all three functions. Three classes of F pili have been found in bacterial cultures: attached, adsorbed, and free. Bacteria with adsorbed F pili in addition to attached ones were proportionately more susceptible to male phage infection, suggesting that adsorbed F pili may be at least partially functional. Free F pili did not compete with bacteria for phage. Some implications of the virus-like nature of F-pilus outgrowth for the mechanisms of mating and male phage infection are discussed.  相似文献   

3.
Some Effects of Temperature on the Growth of F Pili   总被引:19,自引:12,他引:7  
The effect of temperature on the production of F pili by an F(+) strain of Escherichia coli B/r was studied by electron microscopy and by a technique involving serum-blocking power. The latter method is based on the ability of F pili to adsorb F pili antibody which inhibits male-specific phage infection. The total amount of pili in a sample was estimated by serum-blocking power; the length of F pili and number per cell was determined by electron microscopy. Cell extracts prepared by sonic oscillation lacked serum-blocking power, suggesting that F pili are not present in the cytoplasm. The number of F pili per cell varied with the growth temperature, but the average length of F pili remained constant. Maximum number of pili per cell occurs between 37 and 42 C; below 37 C the number decreases, reaching zero at about 25 C. When cells are grown at 37 C, blended, and resuspended in fresh media at 25 C, they make F pili. These pili are probably assembled from a pool of subunits that were synthesized during growth at 37 C. The rates of assembly at 25 and 37 C, as judged by the rate of increase in length of F pili, are similar. When cells were grown at 25 C and shifted up to 37 C, there was a 30-min lag in pili production followed by a period of rapid outgrowth. When cells were shifted down from 37 to 20 C, outgrowth (assembly) of pili ceased, and approximately 50% of the attached pili were released in 2 min. No release was observed when cells were shifted to 0 C. This suggests that pili may be released from the cell by a mechanism that requires metabolic activity, but not the outgrowth of F pili.  相似文献   

4.
An F lac pro mutant which was temperature sensitive for infection by the filamentous bacteriophage f1 but resistant to the F-specific icosahedral RNA phage f2 was isolated. Cells carrying the F' mutation failed to elaborate F pili at all temperatures. Mutant cells were able to pair with recipient cells during bacterial conjugation, but transfer of conjugal DNA occurred at a greatly reduced frequency. Complementation analyses showed the F' mutation to be in the traC gene. When a plasmid carrying traC was introduced into hosts harboring the F' mutation, phage sensitivity, the ability to elaborate F pili, and conjugation efficiency were restored. The mutation was named traC1044. The F lac pro traC1044 mutant appears to be unique among traC mutants in retaining host sensitivity to the filamentous phage f1 in the absence of expression of extended F pili. Phage f1 attachment sites appeared to be present at the cell surface in traC1044 mutants. The reduced accessibility of these sites may account for the reduced efficiency of phage f1 infection of traC1044 hosts, although the possibility that a defect was present in the receptor site itself was not eliminated. Membranes of hosts carrying the F' mutation contained a full complement of mature F-pilin subunits, so the product of traC is presumably required for pilus assembly but not for pilin processing. This, together with the deficiency in conjugal DNA transfer, suggests that traC may be part of a membrane-spanning tra protein complex responsible for pilus assembly and disassembly and conjugal DNA transmission.  相似文献   

5.
Evidence for Two States of F Pili   总被引:1,自引:1,他引:0       下载免费PDF全文
The addition of phenethyl alcohol (PEA) to cultures of male strains of Escherichia coli rapidly prevents the adsorption of the male-specific bacteriophages f1 and f2 to the donor cells. The adsorption of f2 to F pili in cell-free preparations is unaffected by PEA. In a mating system, PEA alters the kinetics of gene transfer in minimal medium but not in broth. Sodium cyanide, azide, and iodoacetate also apparently inhibit f2 adsorption to cells but not to detached F pili. The phage adsorption inhibitory action of PEA is completely reversible in the presence of 100 mug of chloramphenicol per ml.  相似文献   

6.
Coordination of Sex Pili with their Specifying R Factors   总被引:2,自引:0,他引:2  
A single bacterial cell can simultaneously carry both F-like (fi+) and I-like (fi?) R factors and, when the R factors are de-repressed, most cells produce both F-like and I-like sex pili. These pili can be distinguished immunologically and by their capacity to adsorb different phages1. The F pilus is the receptor for RNA phages such as MS2 and filamentous DNA phages such as M13. The I pilus is the receptor for other filamentous DNA phages such as If1 and If2. Electron microscopy suggests that these filamentous DNA phages, both F-specific and I-specific, adsorb to the tip of the pilus2,3.  相似文献   

7.
Role of F Pili in the Penetration of Bacteriophage fl   总被引:28,自引:3,他引:25  
Early stages of infection of Escherichia coli with the filamentous bacteriophage f1 were examined in the electron microscope. Purified phage-bacteria complexes were prepared at various time intervals after the initiation of synchronous infection. Cells were scored for the total number of F pili, the number of F pili with f1 attached, the number of intact phage particles which occurred at the surface of the cell, and F pilus length. Electron microscope autoradiographs were also prepared at each time interval. The results showed that the average number of F pili with f1 attached decreased with time as phage deoxyribonucleic acid (DNA) entered the cell. Concomitant with this loss, the remaining F pili became shorter. The rate of entry of phage DNA into the cell followed, with a short lag, the rate of loss of F pili with f1 attached. During the lag period, intact phage particles accumulated at the surface of the cell. The results from radioautographs showed that no phage DNA could be located within the F pilus. These results suggest that F pili are resorbed by the cell during infection with the bacteriophage f1. Parallel experiments with noninfected cultures further suggest that pilus resorption may be a normal cellular phenomenon.  相似文献   

8.
Evidence is presented for the specific role of sex pili in the eliminatory action of sodium dodecyl sulfate (SDS) on sex (F) and drug resistance (R) factors in Escherichia coli K-12 strains leading to their loss. SDS at 0.03% concentration lysed JE3100 F(8) (+) (F-gal)/gal(-)fla(-)pil(-) in Penassay broth after they had grown exponentially and reached maximum growth to the extent that the agent at concentrations higher than 1% did. However, the agent was only effective in eliminating sex factors from JE3100 in high frequencies at concentrations higher than 1%. Increase of osmotic pressure of the culture with SDS at concentrations as low as 0.03 to 0.1% by addition of sucrose led to the substantial increase of elimination efficiency. Reconstruction experiments between F(8) (+) and F(-) cells in the SDS culture revealed the selective growth of F(-) cells as well as a delay of maximum growth of F(-) variants derived from F(8) (+) cells, compared with those of F(8) (+) cells, as well as F(-) cells originally added to the culture. The agent was not very effective in eliminating sex factors from JE3427 F(8)m(+)5/fla(-)pil(-) cells which lack the function of production of F pili. F(8)m(+)5 cells showed a sensitivity toward SDS intermediate between those of F(8) (+) and F(-) cells. SDS was further effective in eliminating R factors from KE132 R(100-1) (+)/fla(-)pil(-) cells in high efficiency; however, the action was not efficient with KE133 F(100) (+) cells possibly with fewer sex pili than R(100-1) (+). Action of acridine orange on these F(+) or R(+) strains was found to be different in some aspects from that of SDS.  相似文献   

9.
V N Gorelov  T S Il'ina  G B Smirnov 《Genetika》1979,15(7):1206-1220
Assuming the similarity of the processes of illegitimate recombination, such as deletion formation, with the process of F' plasmid formation, we have undertaken the study of the influence of recA- and seg- alleles of Hfr donor on the F' plasmid formation. The data obtained demonstrate the strong influence of donor genotype on the frequency of F' plasmid formation and on the nature of F' plasmids formed, thus demonstrating that the most of F' plasmids have been formed via recombination in Hfr donor cells. The recA- mutation decreased the total yield of F' plasmids selected using both proximal and distal Hfr markers and affected drastically the distribution of the F' plasmids inheriting different proximal unselected markers. The existence of recA-dependent and recA-independent modes of F' plasmid formation was demonstrated. The Escherichia coli chromosome contains regions which involve preferentially in recA-dependent (between proA and gal, and clockwise from gal) or recA-independent (between leu and proA, and the region counterclockwise from argE) recombination. The seg-2 mutation causes only partial block of both recA-dependent and recA-independent recombination pathways, however it causes dramatic decrease of genetic exchanges leading to the formation of the type II F' plasmids. Both seg- and recA- mutations decrease the frequency of the formation of Tra+ F' transconjugants. The percent of Tra- transconjugants, which remain sensitive to MS2 and Q beta donor specific phages, also drops significantly under the influence of the recA- and seg- alleles. Thus, the recombination involving the F structure in wild type strains and seg- mutants occures preferentially in the points of F outside the regions essential for transfer and sensitivity to male specific phages, while in recA- and recA-ges- strains the points inside these regions (tra operon) frequently involved in F' plasmid looping out. There exist more strict correlation between the fertility and sensitivity to phage Q beta than to phage MS2.  相似文献   

10.
Nearly all of 62 strains of Salmonella paratyphi B were sensitive to colicin M and phage T5 but resistant to phages T1 and ES18 and to colicin B. All tested S. typhimurium strains were resistant to colicin M and phage T5, and many were sensitive to phage ES18. A rough S. typhimurium LT2 strain given the tonA region of Escherichia coli or S. paratyphi B became sensitive to colicin M and phage T5. We infer that the tonA allele of S. paratyphi B, like that of E. coli, determines an outer membrane protein that adsorbs T5 and colicin M but not phage ES18, whereas the S. typhimurium allele determines a protein able to adsorb only ES18. The partial T1 sensitivity of a rough LT2 strain with a tonA allele from E. coli or S. paratyphi B and also the tonB(+) phentotype of an E. coli B trp-tonB Delta mutant carrying an F' trp of LT2 origin showed that S. typhimurium LT2 has a tonB allele like that of E. coli with respect to determination of sensitivity to colicins and phage T1. Rough S. paratyphi B, although T5 sensitive, remained resistant to T1 even when given F' tonB(+) of E. coli origin. Classes of Salmonella mutants selected as resistant to colicin M included: T5-resistant mutants, probably tonA(-); mutants unchanged except for M resistance, perhaps tolerant; and Exb(+) mutants, producing a colicin inhibitor (presumably enterochelin). Some Exb(+) mutants were resistant to a bacteriocin inactive on E. coli but active on all tested S. paratyphi B and S. typhimurium strains (and on nearly all other tested Salmonella). A survey showed sensitivity to colicin M in several other species of Salmonella.  相似文献   

11.
The effects of high temperatures (46 to 50 degrees C) on the production of F pili by Escherichia coli were studied by electron microscopy. Attached F pili rapidly disappeared at 48 and 50 degrees C but not at 46 degrees C. Free pili were not denatured at these temperatures. The pili that disappeared from the cells at 50 degrees C did not appear as free pili in the culture supernatant fluid, indicating that the pili had retracted to the cell surface or into the cell. The adsorption of either R17 phage or F pili antibody to the sides of pili prevented retraction. The disappearance of pili was accompanied by a loss in the ability to adsorb R17 phage but not M13 phage, suggesting that the tip of a pilus remains exposed after retraction.  相似文献   

12.
A method for the enumeration of male-specific bacteriophages in sewage   总被引:13,自引:0,他引:13  
Male-specific bacteriophages adsorb to F-pili and thus can only infect male host strains. A method was developed for the selective enumeration of these phages, based on the observation that in sewage there are few phages capable of infecting F- -salmonellas--usually less than 10 pfu/ml. Using a male Salmonella strain, constructed by the introduction of the plasmid F'42 lac::Tn5 into Salmonella typhimurium phage type 3, plaque counts in secondary effluent were found to be in the range of 60-8200 pfu/ml. Practically all the phages detected had a host range restricted to male Salmonella or Escherichia coli strains, were resistant to chloroform and their infectivity was inhibited by RNase. Electron microscopy of lysates revealed phage particles that were morphologically identical to the male-specific single-strand RNA phages. Similar results were obtained with a strain of Salm. indiana carrying F'42 lac. A derivative of the Salm. typhimurium LT2 strain carrying an F-plasmid (F'42 lac fin P301) derepressed for fertility inhibition by the resident plasmid pSLT was equally sensitive to male-specific phages, but from sewage samples many other phages infecting F- E. coli but not F- Salmonella were isolated using this host strain.  相似文献   

13.
When either the F' lac or the F'Cm plasmid was transferred from Escherichia coli into Pasteurella pseudotuberculosis, the P. pseudotuberculosis (F') strains isolated formed plaques with both ribonucleic acid (RNA)-containing and deoxyribonucleic acid-containing male-specific phages. In contrast, strains of P. pestis harboring E. coli (F') plasmids did not form plaques with male-specific phages, although such strains permitted limited multiplication of phage MS2. The adsorption and burst size of MS2 were approximately the same in both species of Pasteurella, but the per cent of adsorbed MS2 that produced infective centers was much lower in P. pestis than it was in P. pseudotuberculosis. By use of a sib-selection technique of P. pestis (F') cells, we isolated a single clone that could form MS2 plaques. (32)P-labeled MS2 adsorbed equally to and its RNA penetrated equally into both the typical MS2-nonpermissive P. pestis cells and the MS2-permissive P. pestis cells. No host modification occurred after growth of MS2 in Pasteurella. Our data suggest that typical strains of P. pestis inhibit the intracellular development of phage MS2.  相似文献   

14.
The bacterial drug resistance plasmids R711b and R778b, at present classified in the X incompatibility group, determine pili (designated 711) that resemble F pili morphologically. Like F pili, 711 pili adsorb F-specific filamentous bacteriophages to their tips, though more often in pairs, than singly. However, F-specific RNA-containing bacteriophages are not adsorbed to their sides, and strains carrying the plasmids are resistant to these phages. Pili determined by the only IncFV plasmid Folac are similar to 711 pili in their phage adsorption properties, but they are serologically different, as are F pili. It is concluded that F, Folac and 711 pili have basic differences in spite of a morphological resemblance.  相似文献   

15.
Effect of Thymine-5-Bromouracil Substitution on F Pili   总被引:4,自引:3,他引:1       下载免费PDF全文
The effect of thymine-5-bromouracil substitution on the regeneration and length of F pili produced by an F(+)Lac(+)/Lac(-)Thy(-) strain of Escherichia coli was studied by electron microscopy. When 5-bromouracil (5BU) incorporation into deoxyribonucleic acid (DNA) was maximal, the modal length of the pilus doubled and the number of pili per cell was approximately 50% that of thymine-grown cells. The ability of 5BU-grown cells to form mating pairs and to be infected by ribonucleic acid (R17) and DNA (M13) male-specific phages was also reduced by approximately 50%. Loss of function was not due to loss of sex factor as 5BU cells retained a sex factor that was susceptible to curing by acridine orange. Elongation of pili on 5BU-grown cells was more sensitive to irradiation at 253.7 nm than on thymine-grown cells, suggesting that DNA is the sensitive target.  相似文献   

16.
Mutants resistant to the donor-specific bacteriophage R17 were isolated from Hfr and Flac-containing strains of Escherichia coli K-12. Thirty-five mutants were examined for the presence of F pili by electron microscopy. The pilus morphology was studied, as were the abilities of the cells to retract their pili and to synthesize new pili. Measurements were made of the efficiency of the conjugal deoxyribonucleic acid transfer and of M13 and R17 phage infection. All mutants had noticeable defects in pilus production, structure, or function. Mutants were found which produced unusually long pili, displayed wide variations in the number of pili per cell, and were deficient in pilus retraction and synthesis. Evidence is presented that there may be two pathways of pilus retraction.  相似文献   

17.
A method for the enumeration of male-specific bacteriophages in sewage   总被引:3,自引:3,他引:0  
H avelaar , A.H. & H ogeboom , W.M. 1984. A method for the enumeration of male-specific bacteriophages in sewage. Journal of Applied Bacteriology 56 , 439–447.
Male-specific bacteriophages adsorb to F-pili and thus can only infect male host strains. A method was developed for the selective enumeration of these phages, based on the observation that in sewage there are few phages capable of infecting F--salmonellas—usually less than 10 pfu/ml. Using a male Salmonella strain, constructed by the introduction of the plasmid F'42 lac::Tn5 into Salmonella typhimu-rium phage type 3, plaque counts in secondary effluent were found to be in the range of 60–8200 pfu/ml. Practically all the phages detected had a host range restricted to male Salmonella or Escherichia coli strains, were resistant to chloroform and their infectivity was inhibited by RNase. Electron microscopy of lysates revealed phage particles that were morphologically identical to the male-specific single-strand RNA phages. Similar results were obtained with a strain of Salm. indiona carrying F'42 lac . A derivative of the Salm. typhimurium LT2 strain carrying an F-plasmid (F'42 lac fin P301) derepressed for fertility inhibition by the resident plasmid pSLT was equally sensitive to male-specific phages, but from sewage samples many other phages infecting F- E. coli but not F- Salmonella were isolated using this host strain.  相似文献   

18.
Previously we have described a mutant Hfr strain in which incompatibility between the integrated F factor and an autonomous F-prime (F') factor was abolished. The mutation (inc) was located in the integrated F factor. F-prime factors isolated from the mutant Hfr strain have the same incompatibility behavior as those isolated from normal Hfr strains. Reintegration of these F' factors into the chromosome restores the Inc- phenotype characteristic of the mutant Hfr. The inc mutation thus affects incompatibility between integrated F and autonomous F(Fi-Fa incompatibility) but not incompatibility between two autonomous F factors (Fa-Fa incompatibility). The implications of this finding for the mechanism of plasmid incompatibility are discussed.  相似文献   

19.
The plasmids R711b (at present IncX) and F0lac (IncFV) both determine pili morphologically like those of F (IncFI), and confer sensitivity to the F-specific filamentous bacteriophages, but not to the F-specific isometric RNA phages. Detailed serological studies show that the two pilus types are unrelated, and that neither is related to any of the previously defined F pilus serotypes. Adsorption of the isometric RNA phage MS2 to R711b pili occurs in the presence but not in the absence of formalin, which presumably prevents elution of reversibly adsorbed virions. No adsorption occurs with F0lac pili. MS2 multiplication, as measured by titre increase tests in liquid medium, is found with neither plasmid. The two plasmids are not incompatible. These observations indicate that R744b and F0lac are different both from one another and from the plasmids belonging to the incompatibility groups IncFI--IV.  相似文献   

20.
A Salmonella typhimurium strain was given the amber mutation hisC527 by transduction, made galactose-negative by mutation, then infected with the F'-1-gal factor. Of 107 spontaneous and mutagen-induced histidine-independent mutants tested, 3 proved to result from suppressor mutations within the F' factor. The mutant F' factors, when transferred to S. typhimurium and E. coli auxotrophs, suppressed amber and ochre but not UGA or missense mutants, and are inferred to carry ochre suppressor genes. Attempts to isolate an F' amber suppressor mutant were unsuccessful. A suppressor F' factor was transferred to 14 rough mutants which had been isolated from LT2 hisC527 (amber) by selection for resistance to phage P22.c2. One rough mutant was partly suppressed, as shown by its acquisition of O agglutinability and by alterations in its phage resistance pattern. Phage P22h grown on the suppressed mutant contransduced its rf. gene with cysE(+) and with pyrE(+), and the affected locus is inferred to be rfaL. Both the original and the mutant F' factors conferred resistance to the rough-specific phage Br60, which is therefore "female-specific."  相似文献   

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