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

Thompson, Russell; Achtman, Mark

doi:10.1007/bf00332530

Cited by 95 publications

(42 citation statements)

References 26 publications

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“…The positive reaction was determined as a macroscopically visible aggregation of bacteria appeared within 60 s. Protein expression in minicells Plasmid pPAP5 and its derivatives were transformed into the minicellproducing strain P678-54 (Adler et al, 1967). Preparation and labelling of plasmid containing minicells with [35S]methionine were as described by Thompson and Achtman (1978). The radioactive samples were subjected to SDS-polyacrylamide electrophoresis (Laemmli, 1970).…”

Section: Chemicals and Enzymesmentioning

confidence: 99%

Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells.

Lindberg¹,

Lund²,

Normark³

1984

The EMBO Journal

189

175

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Most pyelonephritic Escherichia coli strains bind to digalactoside‐containing glycolipids on uroepithelial cells. Purified Pap pili (pili associated with pyelonephritis) show the same binding specificity. A non‐polar mutation early in the papA pilin gene abolishes formation of Pap pili but does not affect the degree of digalactoside‐specific hemagglutination. Three novel pap genes, papE, papF and papG are defined in this report. The papF and papG gene products are both required for digalactoside‐specific agglutination by whole bacteria cells as well as for agglutination by pilus preparations. Pili prepared from a papE mutant have lost their binding ability although whole cells from this mutant retain it, implying an adhesin anchoring role for the papE gene product. A mutant with lesions both in the papA and the papE genes does not mediate digalactoside‐specific agglutination. The implications of this finding for pilus biogenesis are discussed.

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Section: Chemicals and Enzymesmentioning

confidence: 99%

Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells.

Lindberg¹,

Lund²,

Normark³

1984

The EMBO Journal

189

175

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“…Plasmids were transformed into ORN103, and minicells were prepared and labeled with 80 ,uCi of [35S]methionine as described previously (41). Immunoprecipitation was performed as previously outlined (35).…”

mentioning

confidence: 99%

PapD, a periplasmic transport protein in P-pilus biogenesis

et al. 1989

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The product of the papD gene of uropathogenic Escherichia coli is required for the biogenesis of digalactoside-binding P pili. Mutations within papD result in complete degradation of the major pilus subunit, PapA, and of the pilinlike proteins PapE and PapF and also cause partial breakdown of the PapG adhesin. The papD gene was sequenced, and the gene product was purified from the periplasm. The deduced amino acid sequence and the N-terminal sequence obtained from the purified protein revealed that PapD is a basic and hydrophilic peripheral protein. A periplasmic complex between PapD and PapE was purified from cells that overproduced and accumulated these proteins in the periplasm. Antibodies raised against this complex reacted with purified wild-type P pili but not with pili purified from a papE mutant. In contrast, anti-PapD serum did not react with purified pili or with the culture fluid of piliated cells. However, this serum was able to specifically precipitate the PapE protein from periplasmic extracts, confirming that PapD and PapE were associated as a complex. It is suggested that PapD functions in P-pilus biogenesis as a periplasmic transport protein. Probably PapD forms complexes with pilus subunits at the outer surface of the inner membrane and transports them in a stable configuration across the periplasmic space before delivering them to the site(s) of pilus polymerization.

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“…The three inverted repeats in this region are also shown; the third set is the proposed terminator for traM transcription (17). BglII digests of plasmids encoding type I or type II finP genes were ligated into pUC8 (19), and chimera-containing colonies were immobilized on nitrocellulose and probed with a 32P-labeled DNA probe (9) made from the 1.1-kilobase BglII fragment of F that contains oriT, traM, andfinP (16,17). Plasmid DNA was prepared (1) from colonies containing the 1.1-kilobase BglII fragment and sequenced by the method of Wallace et al (20), using reverse or 17-bp primers (New England BioLabs, Inc.).…”

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confidence: 99%

Nucleotide sequences of five IncF plasmid finP alleles

Finlay¹,

Frost²,

Paranchych³

et al. 1986

J Bacteriol

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The nucleotide sequences of fivefinP alleles from various IncF plasmids (finP types I to V) as well as of three finP mutations were determined and compared. ThefinP gene specificity could be attributed to a variable, sixto-seven-nucleotide loop located between inverted repeats, and the sequence data were consistent with the product offinP being an RNA molecule rather than a protein. ThefinP mutations interrupted a proposedfinP promoter or destabilized a predicted stem-and-loop structure in the finP RNA molecule.Conjugal DNA transfer requires expression of the transfer (tra) operon which is regulated in a two-stage process involving the fertility inhibition system (finO and finP) and the traJ gene (4, 5). The products offinO and finP (FinO and FinP) are regulatory elements that act together at the fisO site (otherwise designated Oj or traO) to prevent traJ transcription. The traJ gene product, a positive control element, interacts with the pyz promoter, increasing transcription of the tra YZ operon (6,11,21).All finO products from IncF plasmids studied to date are capable of complementing the naturalfinO mutation of the F plasmid (22) and inhibiting its transfer. There may be two types offinO products, differentiated by their various abilities to inhibit this transfer (22). ThefinO genes from plasmids R100 and R6-5 have been cloned, but the finO product remains undefined (2,3,18).The finP gene is located between traM and traJ (8), and the nucleotide sequence of this region from the F plasmid has been determined (17). Mullineaux and Willetts (11) suggested that finP is transcribed from a potential promoter within the translated region of the traJ transcript and that it is transcribed in the opposite direction from traJ transcription.FinP is plasmid specific, since six finP alleles were found among the 12 IncF plasmids studied (22 (11,22). Wild-type finP regions of types II to V (22) from the IncF plasmids R124, ColVBtrp, R1-19, R100-1, and ColB4 were cloned, sequenced, and compared with the sequence of the finP region of the F plasmid (type I) (Fig. 1). In addition, the sequences of three finP mutations, plasmid pED236 (ColB2Fdr, type II), plasmid pED200 (R124 finP, type II), and plasmid pED202 (R386finP, type VI, or R386fisO, type VI, or both), which lead to derepression of the transfer operon in these plasmids were determined (Fig. 1) The finP promoter was located within a BglII-TaqI fragment in F corresponding to positions 1 to 84 in Fig. 1 (11) and may overlap the BamHI site at position 35 in R100-1 (type IV) (B. E. Fee and W. B. Dempsey, submitted for publication). It is evident that the -35 and -10 regions of a putative promoter are highly conserved in all the finP alleles examined (positions 10 to 38). Except for position 15, the -35 region of this finP promoter matches the procaryotic consensus sequence TTGACa (7). The sequence at the proposed -10 region is conserved in four of the five alleles; only R100-1 differs at position 35. This sequence also strongly resembles the proposed consensus sequence for...

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

Cited by 95 publications

References 26 publications

Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells.

Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells.

PapD, a periplasmic transport protein in P-pilus biogenesis

Nucleotide sequences of five IncF plasmid finP alleles

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