Induction of gene expression in Escherichia coli after pilus-mediated adherence

Science

Volumn 273, Issue 5279, 1996, Pages 1234-1236

  Zhang, Jian Ping ^a,c   Normark, Staffan ^a,b  

^a WASHINGTON UNIVERSITY SCHOOL OF MEDICINE   (United States)

^b KAROLINSKA INSTITUTE   (Sweden)

^c Microcide Pharmaceuticals   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

IRON; MEMBRANE RECEPTOR; SIDEROPHORE; VIRULENCE FACTOR;

ANIMAL CELL; ARTICLE; BACTERIAL GROWTH; BACTERIUM ADHERENCE; BACTERIUM PILUS; CELL CONTACT; CONTROLLED STUDY; ERYTHROCYTE; ESCHERICHIA COLI; GENE EXPRESSION; HOST CELL; HUMAN; HUMAN CELL; NONHUMAN; PRIORITY JOURNAL; RABBIT; REGULATOR GENE; URINARY TRACT INFECTION;

ANIMALIA; BACTERIA (MICROORGANISMS); ESCHERICHIA COLI; ORYCTOLAGUS CUNICULUS;

EID: 0000737074     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5279.1234     Document Type: Article

References (29)

1. J. R. Johnson, Clin. Microbiol. Rev. 4, 80 (1991).
2. J. A. Roberts et al., Proc. Natl. Acad. Sci. U.S.A 91, 11889 (1994).
3. M. J. Kuehn et al., Nature 356, 252 (1992); E. Bullitt and L. Makowski, ibid. 373, 164 (1995).
4. M. J. Kuehn et al., Nature 356, 252 (1992); E. Bullitt and L. Makowski, ibid. 373, 164 (1995).
5. N. Stromberg et al., EMBO J. 9, 2001 (1990).
6. K. K. Wong and M. McClelland, Proc. Natl. Acad. Sci. U.S.A. 91, 639 (1994); P. Liang and A. B. Pardee, Science 257, 967 (1992).
7. K. K. Wong and M. McClelland, Proc. Natl. Acad. Sci. U.S.A. 91, 639 (1994); P. Liang and A. B. Pardee, Science 257, 967 (1992).
8. Attachment and RT-PCR procedures were as follows: DS17 or DS17-8 were grown at 37°C in LB broth overnight, and 1 ml of overnight culture was inoculated into 9 ml of fresh LB broth. Cultures were incubated for 3 hours more, and 0.5 ml of 10% glutaraldehyde-stabilized human group O RBCs or rabbit RBCs were added to the cultures. After 30 min to allow for attachment, cells were pelleted, and the pellets of bacteria and RBCs were lysed in 1 ml of TRIzol total RNA isolation reagent (Gibco BRL). Total RNA isolation was continued according to TRIzol instructions. Approximately 2 mg of total RNA harvested from each sample was reverse-transcribed and then used as templates for PCR with a nonspecific set of primers as described in (5). The PCR parameters were 94°C for 1 min, 42°C for 1 min, and 72°C for 1 min for a total of 40 cycles, followed by 10 min of elongation at 72°C.
9. S. Nagasawa et al., Mol. Microbiol. 6, 799 (1992); K. Ishige et al., EMBO J. 13, 5195 (1994).
10. S. Nagasawa et al., Mol. Microbiol. 6, 799 (1992); K. Ishige et al., EMBO J. 13, 5195 (1994).
11. 4 or CDH dissolved in 3 ml of methanol. The DS17 or DS17-8 strains were grown in LB broth overnight at 37°C, and 1 ml of overnight culture of each was inoculated into 9 ml of fresh LB broth. Bacteria were grown for another 3 hours and then centrifuged. Nine milliliters of culture supernatant was removed, and the bacterial pellets were resuspended in the remaining 1 ml of culture media. The bacterial suspensions were added to the pre-coated petri dishes at 37°C for 30 min with gentle shaking (60 rpm). Bacteria were lysed on the plates with 5 ml of TRIzol solution, and total RNA was isolated as above. Specific airS primers are listed above. The pyruvate dehydrogenase primers were pdh-1 (5′-TTGCTCGAAACACGACACGCCCT-3′) and pdh-2 (5′-ACGGCAATCTGATACTGGAGTAC-3′). PCR products were resolved on 8% polyacrylamide gels.
12. A. H. Free and H. M. Free, in Urinalysis in Clinical Laboratory Practice (CRC Press, Cleveland, 1975), chap. 4, pp. 13-19.
13. J. B. Neilands, Can. J. Microbiol. 38, 728 (1992).
14. M. L. Guerinot, Annu. Rev. Microbiol. 48, 743 (1994).
15. J. A. Robledo et al., J. Urol. 143, 386 (1990).
16. C. D. Nau and J. Konisky, J. Bacteriol. 171, 1041 (1989).
17. B. Arico et al., Proc. Natl. Acad. Sci. U.S.A. 86, 6671 (1989); A. R. Melton and A. A. Weiss, J. Bacteriol. 170, 6206 (1989); P. C. Giardina et al., ibid. 177, 6058 (1995).
18. B. Arico et al., Proc. Natl. Acad. Sci. U.S.A. 86, 6671 (1989); A. R. Melton and A. A. Weiss, J. Bacteriol. 170, 6206 (1989); P. C. Giardina et al., ibid. 177, 6058 (1995).
19. B. Arico et al., Proc. Natl. Acad. Sci. U.S.A. 86, 6671 (1989); A. R. Melton and A. A. Weiss, J. Bacteriol. 170, 6206 (1989); P. C. Giardina et al., ibid. 177, 6058 (1995).
20. C. R. Dean and K. Poole, Mol. Microbiol. 8, 1095 (1993).
21. D.K. Willis et al., Mol. Plant-Microbe Interact. 1, 80 (1990).
22. J. Z. Montgonerie et al., Infect. Immun. 46, 835 (1984).
23. S. M. Opal et al., J. Infect. Disease 161, 794 (1990).
24. E. Griffiths et al., Infect. Immun. 47, 808 (1985).
25. C. M. Hamilton et al., J. Bacteriol. 171, 4617 (1989).
26. Y. Kohara et al., Cell 50, 495 (1987).
27. J. Brosius, Gene 27, 151 (1984).
28. J. R. Neumann et al., Biotechniques 5, 444 (1987).
29. We thank M. Caparon for providing the kanamycin cassette, S. R. Kushner for the plasmid pMAK705, and J. W. St. Gerne for critically reading the manuscript. Supported by NIH grant 5RO1GM44655-05, the Swedish Medical Research Council, the Göran Gustafsson Foundation of Natural and Medical Science, and an unrestrictive grant for infectious disease research from Bristol-Myers Squibb.