Abstract
Copper-zinc superoxide dismutases (Cu,Zn SODs), until recently considered very unusual in bacteria, are now being found in a wide range of gram-negative bacterial species. Here we report the cloning and characterization of sodC, encoding Cu,Zn SOD in Actinobacillus pleuropneumoniae, a major pathogen of pigs and the causative organism of porcine pleuropneumonia. sodC was shown to lie on a monocistronic operon, at the chromosomal locus between the genes asd (encoding aspartate semialdehyde dehydrogenase) and recF. The primary gene product was shown to have an N-terminal peptide extension functioning as a leader peptide, so that the mature Actinobacillus enzyme, like other bacterial examples, is directed to the periplasm, where it is appropriately located to dismutate exogenously generated superoxide. While the role of these secreted bacterial SODs is unknown, we speculate that in A. pleuropneumoniae the enzyme may confer survival advantage by accelerating dismutation of superoxide derived from neutrophils, a central host defense response in the course of porcine infection.
Full Text
The Full Text of this article is available as a PDF (318.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beaman L., Beaman B. L. Monoclonal antibodies demonstrate that superoxide dismutase contributes to protection of Nocardia asteroides within the intact host. Infect Immun. 1990 Sep;58(9):3122–3128. doi: 10.1128/iai.58.9.3122-3128.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beauchamp C., Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971 Nov;44(1):276–287. doi: 10.1016/0003-2697(71)90370-8. [DOI] [PubMed] [Google Scholar]
- Beck B. L., Tabatabai L. B., Mayfield J. E. A protein isolated from Brucella abortus is a Cu-Zn superoxide dismutase. Biochemistry. 1990 Jan 16;29(2):372–376. doi: 10.1021/bi00454a010. [DOI] [PubMed] [Google Scholar]
- Benov L. T., Fridovich I. Escherichia coli expresses a copper- and zinc-containing superoxide dismutase. J Biol Chem. 1994 Oct 14;269(41):25310–25314. [PubMed] [Google Scholar]
- Bilinski T. Oxygen toxicity and microbial evolution. Biosystems. 1991;24(4):305–312. doi: 10.1016/0303-2647(91)90049-q. [DOI] [PubMed] [Google Scholar]
- Bordo D., Djinović K., Bolognesi M. Conserved patterns in the Cu,Zn superoxide dismutase family. J Mol Biol. 1994 May 6;238(3):366–386. doi: 10.1006/jmbi.1994.1298. [DOI] [PubMed] [Google Scholar]
- Broome-Smith J. K., Spratt B. G. A vector for the construction of translational fusions to TEM beta-lactamase and the analysis of protein export signals and membrane protein topology. Gene. 1986;49(3):341–349. doi: 10.1016/0378-1119(86)90370-7. [DOI] [PubMed] [Google Scholar]
- Canvin J., Langford P. R., Wilks K. E., Kroll J. S. Identification of sodC encoding periplasmic [Cu,Zn]-superoxide dismutase in Salmonella. FEMS Microbiol Lett. 1996 Feb 15;136(2):215–220. doi: 10.1111/j.1574-6968.1996.tb08052.x. [DOI] [PubMed] [Google Scholar]
- Crapo J. D., McCord J. M., Fridovich I. Preparation and assay of superoxide dismutases. Methods Enzymol. 1978;53:382–393. doi: 10.1016/s0076-6879(78)53044-9. [DOI] [PubMed] [Google Scholar]
- DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
- Deneer H. G., Potter A. A. Effect of iron restriction on the outer membrane proteins of Actinobacillus (Haemophilus) pleuropneumoniae. Infect Immun. 1989 Mar;57(3):798–804. doi: 10.1128/iai.57.3.798-804.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dom P., Haesebrouck F., Ducatelle R., Charlier G. In vivo association of Actinobacillus pleuropneumoniae serotype 2 with the respiratory epithelium of pigs. Infect Immun. 1994 Apr;62(4):1262–1267. doi: 10.1128/iai.62.4.1262-1267.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dom P., Haesebrouck F., Kamp E. M., Smits M. A. Influence of Actinobacillus pleuropneumoniae serotype 2 and its cytolysins on porcine neutrophil chemiluminescence. Infect Immun. 1992 Oct;60(10):4328–4334. doi: 10.1128/iai.60.10.4328-4334.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dom P., Haesebrouck F., Kamp E. M., Smits M. A. NAD-independent Actinobacillus pleuropneumoniae strains: production of RTX toxins and interactions with porcine phagocytes. Vet Microbiol. 1994 Apr;39(3-4):205–218. doi: 10.1016/0378-1135(94)90158-9. [DOI] [PubMed] [Google Scholar]
- Feldman C., Anderson R., Kanthakumar K., Vargas A., Cole P. J., Wilson R. Oxidant-mediated ciliary dysfunction in human respiratory epithelium. Free Radic Biol Med. 1994 Jul;17(1):1–10. doi: 10.1016/0891-5849(94)90002-7. [DOI] [PubMed] [Google Scholar]
- Frey J., Haldimann A., Nicolet J., Boffini A., Prentki P. Sequence analysis and transcription of the apxI operon (hemolysin I) from Actinobacillus pleuropneumoniae. Gene. 1994 May 3;142(1):97–102. doi: 10.1016/0378-1119(94)90361-1. [DOI] [PubMed] [Google Scholar]
- Frey J. Virulence in Actinobacillus pleuropneumoniae and RTX toxins. Trends Microbiol. 1995 Jul;3(7):257–261. doi: 10.1016/s0966-842x(00)88939-8. [DOI] [PubMed] [Google Scholar]
- Hassan H. M., Sun H. C. Regulatory roles of Fnr, Fur, and Arc in expression of manganese-containing superoxide dismutase in Escherichia coli. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3217–3221. doi: 10.1073/pnas.89.8.3217. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Haziza C., Stragier P., Patte J. C. Nucleotide sequence of the asd gene of Escherichia coli: absence of a typical attenuation signal. EMBO J. 1982;1(3):379–384. doi: 10.1002/j.1460-2075.1982.tb01178.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Imlay K. R., Imlay J. A. Cloning and analysis of sodC, encoding the copper-zinc superoxide dismutase of Escherichia coli. J Bacteriol. 1996 May;178(9):2564–2571. doi: 10.1128/jb.178.9.2564-2571.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inzana T. J. Virulence properties of Actinobacillus pleuropneumoniae. Microb Pathog. 1991 Nov;11(5):305–316. doi: 10.1016/0882-4010(91)90016-4. [DOI] [PubMed] [Google Scholar]
- James E. R., McLean D. C., Jr, Perler F. Molecular cloning of an Onchocerca volvulus extracellular Cu-Zn superoxide dismutase. Infect Immun. 1994 Feb;62(2):713–716. doi: 10.1128/iai.62.2.713-716.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jansen R., Briaire J., Smith H. E., Dom P., Haesebrouck F., Kamp E. M., Gielkens A. L., Smits M. A. Knockout mutants of Actinobacillus pleuropneumoniae serotype 1 that are devoid of RTX toxins do not activate or kill porcine neutrophils. Infect Immun. 1995 Jan;63(1):27–37. doi: 10.1128/iai.63.1.27-37.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kroll J. S., Langford P. R., Loynds B. M. Copper-zinc superoxide dismutase of Haemophilus influenzae and H. parainfluenzae. J Bacteriol. 1991 Dec;173(23):7449–7457. doi: 10.1128/jb.173.23.7449-7457.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kroll J. S., Langford P. R., Saah J. R., Loynds B. M. Molecular and genetic characterization of superoxide dismutase in Haemophilus influenzae type b. Mol Microbiol. 1993 Nov;10(4):839–848. doi: 10.1111/j.1365-2958.1993.tb00954.x. [DOI] [PubMed] [Google Scholar]
- Kroll J. S., Langford P. R., Wilks K. E., Keil A. D. Bacterial [Cu,Zn]-superoxide dismutase: phylogenetically distinct from the eukaryotic enzyme, and not so rare after all! Microbiology. 1995 Sep;141(Pt 9):2271–2279. doi: 10.1099/13500872-141-9-2271. [DOI] [PubMed] [Google Scholar]
- Langford P. R., Loynds B. M., Kroll J. S. Copper-zinc superoxide dismutase in Haemophilus species. J Gen Microbiol. 1992 Mar;138(3):517–522. doi: 10.1099/00221287-138-3-517. [DOI] [PubMed] [Google Scholar]
- Latimer E., Simmers J., Sriranganathan N., Roop R. M., 2nd, Schurig G. G., Boyle S. M. Brucella abortus deficient in copper/zinc superoxide dismutase is virulent in BALB/c mice. Microb Pathog. 1992 Feb;12(2):105–113. doi: 10.1016/0882-4010(92)90113-3. [DOI] [PubMed] [Google Scholar]
- Liggett A. D., Harrison L. R., Farrell R. L. Sequential study of lesion development in experimental haemophilus pleuropneumonia. Res Vet Sci. 1987 Mar;42(2):204–212. [PubMed] [Google Scholar]
- Loynds B. M., Langford P. R., Kroll J. S. recF in Actinobacillus pleuropneumoniae. Nucleic Acids Res. 1992 Feb 11;20(3):615–615. doi: 10.1093/nar/20.3.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mulks M. H., Buysse J. M. A targeted mutagenesis system for Actinobacillus pleuropneumoniae. Gene. 1995 Nov 7;165(1):61–66. doi: 10.1016/0378-1119(95)00528-e. [DOI] [PubMed] [Google Scholar]
- Pugsley A. P. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. doi: 10.1128/mr.57.1.50-108.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raibaud O., Schwartz M. Positive control of transcription initiation in bacteria. Annu Rev Genet. 1984;18:173–206. doi: 10.1146/annurev.ge.18.120184.001133. [DOI] [PubMed] [Google Scholar]
- Rather P. N., Moran C. P., Jr Compartment-specific transcription in Bacillus subtilis: identification of the promoter for gdh. J Bacteriol. 1988 Nov;170(11):5086–5092. doi: 10.1128/jb.170.11.5086-5092.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rodden J. L., Scocca J. J. Purification and properties of cyclic phosphodiesterase: 3'-nucleotidase, a periplasmic enzyme of Haemophilus influenzae. Arch Biochem Biophys. 1972 Dec;153(2):837–844. doi: 10.1016/0003-9861(72)90406-7. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schnell S., Steinman H. M. Function and stationary-phase induction of periplasmic copper-zinc superoxide dismutase and catalase/peroxidase in Caulobacter crescentus. J Bacteriol. 1995 Oct;177(20):5924–5929. doi: 10.1128/jb.177.20.5924-5929.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sebunya T. N., Saunders J. R. Haemophilus pleuropneumoniae infection in swine: a review. J Am Vet Med Assoc. 1983 Jun 15;182(12):1331–1337. [PubMed] [Google Scholar]
- Simurda M. C., van Keulen H., Rekosh D. M., LoVerde P. T. Schistosoma mansoni: identification and analysis of an mRNA and a gene encoding superoxide dismutase (Cu/Zn). Exp Parasitol. 1988 Oct;67(1):73–84. doi: 10.1016/0014-4894(88)90010-0. [DOI] [PubMed] [Google Scholar]
- St John G., Steinman H. M. Periplasmic copper-zinc superoxide dismutase of Legionella pneumophila: role in stationary-phase survival. J Bacteriol. 1996 Mar;178(6):1578–1584. doi: 10.1128/jb.178.6.1578-1584.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steinman H. M. Bacteriocuprein superoxide dismutase of Photobacterium leiognathi. Isolation and sequence of the gene and evidence for a precursor form. J Biol Chem. 1987 Feb 5;262(4):1882–1887. [PubMed] [Google Scholar]
- Steinman H. M. Bacteriocuprein superoxide dismutases in pseudomonads. J Bacteriol. 1985 Jun;162(3):1255–1260. doi: 10.1128/jb.162.3.1255-1260.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Steinman H. M., Ely B. Copper-zinc superoxide dismutase of Caulobacter crescentus: cloning, sequencing, and mapping of the gene and periplasmic location of the enzyme. J Bacteriol. 1990 Jun;172(6):2901–2910. doi: 10.1128/jb.172.6.2901-2910.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stoscheck C. M. Increased uniformity in the response of the coomassie blue G protein assay to different proteins. Anal Biochem. 1990 Jan;184(1):111–116. doi: 10.1016/0003-2697(90)90021-z. [DOI] [PubMed] [Google Scholar]
- Tabatabai L. B., Pugh G. W., Jr Modulation of immune responses in Balb/c mice vaccinated with Brucella abortus Cu-Zn superoxide dismutase synthetic peptide vaccine. Vaccine. 1994 Aug;12(10):919–924. doi: 10.1016/0264-410x(94)90035-3. [DOI] [PubMed] [Google Scholar]
- Tainer J. A., Getzoff E. D., Beem K. M., Richardson J. S., Richardson D. C. Determination and analysis of the 2 A-structure of copper, zinc superoxide dismutase. J Mol Biol. 1982 Sep 15;160(2):181–217. doi: 10.1016/0022-2836(82)90174-7. [DOI] [PubMed] [Google Scholar]
- Tatum F. M., Detilleux P. G., Sacks J. M., Halling S. M. Construction of Cu-Zn superoxide dismutase deletion mutants of Brucella abortus: analysis of survival in vitro in epithelial and phagocytic cells and in vivo in mice. Infect Immun. 1992 Jul;60(7):2863–2869. doi: 10.1128/iai.60.7.2863-2869.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson R., Achtman M. The control region of the F sex factor DNA transfer cistrons: restriction mapping and DNA cloning. Mol Gen Genet. 1978 Oct 24;165(3):295–304. doi: 10.1007/BF00332530. [DOI] [PubMed] [Google Scholar]
- Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]