Purification and Characterization of Thin Pili of IncI1 Plasmids ColIb-P9 and R64: Formation of PilV-specific Cell Aggregates by Type IV Pili

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1998 Jun 6

PMID 9603870

Citations 29

Authors

T Yoshida

N Furuya

M Ishikura

T Isobe

T Ogawa

T Komano

Affiliations

Soon will be listed here.

Abstract

Thin pili of the closely related IncI1 plasmids ColIb-P9 and R64 are required only for liquid mating and belong to the type IV family of pili. They were sedimented by ultracentrifugation from culture medium in which Escherichia coli cells harboring ColIb-P9- or R64-derived plasmids had been grown, and then the pili were purified by CsCl density gradient centrifugation. In negatively stained thin pilus samples, long rods with a diameter of 6 nm, characteristic of type IV pili, were observed under an electron microscope. Gel electrophoretic analysis of purified ColIb-P9 thin pili indicated that thin pili consist of two kinds of proteins, pilin and the PilV protein. Pilin was demonstrated to be the product of the pilS gene. Pilin was first synthesized as a 22-kDa prepilin from the pilS gene and subsequently processed to a 19-kDa protein by the function of the pilU product. The N-terminal amino group of the processed protein was shown to be modified. The C-terminal segments of the pilV products vary among six or seven different types, as a result of shufflon DNA rearrangements of the pilV gene. These PilV proteins were revealed to comprise a minor component of thin pili. Formation of PilV-specific cell aggregates by ColIb-P9 and R64 thin pili was demonstrated and may play an important role in liquid mating.

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References

Ottow J . Ecology, physiology, and genetics of fimbriae and pili. Annu Rev Microbiol. 1975; 29:79-108. DOI: 10.1146/annurev.mi.29.100175.000455. View

Kim S, Komano T . The plasmid R64 thin pilus identified as a type IV pilus. J Bacteriol. 1997; 179(11):3594-603. PMC: 179153. DOI: 10.1128/jb.179.11.3594-3603.1997. View

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

Folkhard W, Marvin D, Watts T, Paranchych W . Structure of polar pili from Pseudomonas aeruginosa strains K and O. J Mol Biol. 1981; 149(1):79-93. DOI: 10.1016/0022-2836(81)90261-8. View

BRADLEY D . Derepressed plasmids of incompatibility group I1 determine two different morphological forms of pilus. Plasmid. 1983; 9(3):331-4. DOI: 10.1016/0147-619x(83)90011-2. View

Watts T, Kay C, Paranchych W . Spectral properties of three quaternary arrangements of Pseudomonas pilin. Biochemistry. 1983; 22(15):3640-6. DOI: 10.1021/bi00284a016. View

BRADLEY D . Characteristics and function of thick and thin conjugative pili determined by transfer-derepressed plasmids of incompatibility groups I1, I2, I5, B, K and Z. J Gen Microbiol. 1984; 130(6):1489-502. DOI: 10.1099/00221287-130-6-1489. View

Vieira J, Messing J . Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985; 33(1):103-19. DOI: 10.1016/0378-1119(85)90120-9. View

Komano T, Kubo A, Nisioka T . Shufflon: multi-inversion of four contiguous DNA segments of plasmid R64 creates seven different open reading frames. Nucleic Acids Res. 1987; 15(3):1165-72. PMC: 340515. DOI: 10.1093/nar/15.3.1165. View

10.

Lindberg F, Lund B, Johansson L, Normark S . Localization of the receptor-binding protein adhesin at the tip of the bacterial pilus. Nature. 1987; 328(6125):84-7. DOI: 10.1038/328084a0. View

11.

Bressan G, Stanley K . pUEX, a bacterial expression vector related to pEX with universal host specificity. Nucleic Acids Res. 1987; 15(23):10056. PMC: 306560. DOI: 10.1093/nar/15.23.10056. View

12.

Vieira J, Messing J . Production of single-stranded plasmid DNA. Methods Enzymol. 1987; 153:3-11. DOI: 10.1016/0076-6879(87)53044-0. View

13.

Kubo A, KUSUKAWA A, Komano T . Nucleotide sequence of the rci gene encoding shufflon-specific DNA recombinase in the IncI1 plasmid R64: homology to the site-specific recombinases of integrase family. Mol Gen Genet. 1988; 213(1):30-5. DOI: 10.1007/BF00333394. View

14.

Kim S, Komano T . Cloning and nucleotide sequence of the ColIb shufflon. Plasmid. 1989; 22(2):180-4. DOI: 10.1016/0147-619x(89)90029-2. View

15.

Komano T, Funayama N, Kim S, Nisioka T . Transfer region of IncI1 plasmid R64 and role of shufflon in R64 transfer. J Bacteriol. 1990; 172(5):2230-5. PMC: 208852. DOI: 10.1128/jb.172.5.2230-2235.1990. View

16.

Kuehn M, Heuser J, Normark S, Hultgren S . P pili in uropathogenic E. coli are composite fibres with distinct fibrillar adhesive tips. Nature. 1992; 356(6366):252-5. DOI: 10.1038/356252a0. View

17.

Pugsley A . The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993; 57(1):50-108. PMC: 372901. DOI: 10.1128/mr.57.1.50-108.1993. View

18.

Hultgren S, Abraham S, Caparon M, Falk P, St Geme 3rd J, Normark S . Pilus and nonpilus bacterial adhesins: assembly and function in cell recognition. Cell. 1993; 73(5):887-901. DOI: 10.1016/0092-8674(93)90269-v. View

19.

Kim S, Funayama N, Komano T . Nucleotide sequence and characterization of the traABCD region of IncI1 plasmid R64. J Bacteriol. 1993; 175(16):5035-42. PMC: 204969. DOI: 10.1128/jb.175.16.5035-5042.1993. View

20.

Strom M, Lory S . Structure-function and biogenesis of the type IV pili. Annu Rev Microbiol. 1993; 47:565-96. DOI: 10.1146/annurev.mi.47.100193.003025. View