Abstract
Staphylococcus aureus ATCC 49775 produces three proteins recognized by affinity-purified antibodies against the S component of Panton-Valentine leucocidin (LukS-PV) and two proteins recognized by affinity-purified antibodies against the F component of this toxin (LukF-PV). Purification of these proteins and cloning of the corresponding genes provided evidence for the presence of two loci. The first one, encoding Panton-Valentine leucocidin, consisted of two cotranscribed open reading frames, lukS-PV and lukF-PV, coding the class S and the class F components, respectively. The second one coded for a gamma-hemolysin and consisted of two transcription units, the first one encoding an HlgA-like protein, a class S component, and the second one encoding two cotranscribed open reading frames identical to HlgC and HlgB, class S and class F components, respectively, from gamma-hemolysin from the reference strain Smith 5R. It appears that the Panton-Valentine leucocidin from S. aureus ATCC 49775 (V8 strain) should not be confused with leucocidin from ATCC 27733 (another isolate of V8 strain), which had 95% identity with HlgC and HlgB from gamma-hemolysin. The cosecretion of these five proteins led to six possible synergistic combinations between F and S components. Two of these combinations (LukS-PV-LukF-PV and HlgA-LukF-PV) had dermonecrotic activity on rabbit skin, and all six were leukocytolytic on glass-adsorbed leukocytes. Only three were hemolytic on rabbit erythrocytes, the two gamma-hemolysin combinations and the combination LukF-PV-HlgA.
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- Choorit W., Kaneko J., Muramoto K., Kamio Y. Existence of a new protein component with the same function as the LukF component of leukocidin or gamma-hemolysin and its gene in Staphylococcus aureus P83. FEBS Lett. 1995 Jan 9;357(3):260–264. doi: 10.1016/0014-5793(94)01372-8. [DOI] [PubMed] [Google Scholar]
- Clyne M., Birkbeck T. H., Arbuthnott J. P. Characterization of staphylococcal gamma-lysin. J Gen Microbiol. 1992 May;138(5):923–930. doi: 10.1099/00221287-138-5-923. [DOI] [PubMed] [Google Scholar]
- Colin D. A., Mazurier I., Sire S., Finck-Barbançon V. Interaction of the two components of leukocidin from Staphylococcus aureus with human polymorphonuclear leukocyte membranes: sequential binding and subsequent activation. Infect Immun. 1994 Aug;62(8):3184–3188. doi: 10.1128/iai.62.8.3184-3188.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cooney J., Kienle Z., Foster T. J., O'Toole P. W. The gamma-hemolysin locus of Staphylococcus aureus comprises three linked genes, two of which are identical to the genes for the F and S components of leukocidin. Infect Immun. 1993 Feb;61(2):768–771. doi: 10.1128/iai.61.2.768-771.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Couppie P., Cribier B., Prévost G. Leukocidin from Staphylococcus aureus and cutaneous infections: an epidemiologic study. Arch Dermatol. 1994 Sep;130(9):1208–1209. doi: 10.1001/archderm.130.9.1208. [DOI] [PubMed] [Google Scholar]
- Cribier B., Prévost G., Couppie P., Finck-Barbançon V., Grosshans E., Piémont Y. Staphylococcus aureus leukocidin: a new virulence factor in cutaneous infections? An epidemiological and experimental study. Dermatology. 1992;185(3):175–180. doi: 10.1159/000247443. [DOI] [PubMed] [Google Scholar]
- Finck-Barbançon V., Duportail G., Meunier O., Colin D. A. Pore formation by a two-component leukocidin from Staphylococcus aureus within the membrane of human polymorphonuclear leukocytes. Biochim Biophys Acta. 1993 Oct 20;1182(3):275–282. doi: 10.1016/0925-4439(93)90069-d. [DOI] [PubMed] [Google Scholar]
- Finck-Barbançon V., Prévost G., Piémont Y. Improved purification of leukocidin from Staphylococcus aureus and toxin distribution among hospital strains. Res Microbiol. 1991 Jan;142(1):75–85. doi: 10.1016/0923-2508(91)90099-v. [DOI] [PubMed] [Google Scholar]
- GLADSTONE G. P., VAN HEYNINGEN W. E. Staphylococcal leucocidins. Br J Exp Pathol. 1957 Apr;38(2):123–137. [PMC free article] [PubMed] [Google Scholar]
- Guyonnet F., Plommet M. Hémolysine gamma de staphylococcus aureus: purification et propriétés. Ann Inst Pasteur (Paris) 1970 Jan;118(1):19–33. [PubMed] [Google Scholar]
- Hensler T., König B., Prévost G., Piémont Y., Köller M., König W. Leukotriene B4 generation and DNA fragmentation induced by leukocidin from Staphylococcus aureus: protective role of granulocyte-macrophage colony-stimulating factor (GM-CSF) and G-CSF for human neutrophils. Infect Immun. 1994 Jun;62(6):2529–2535. doi: 10.1128/iai.62.6.2529-2535.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kamio Y., Rahman A., Nariya H., Ozawa T., Izaki K. The two Staphylococcal bi-component toxins, leukocidin and gamma-hemolysin, share one component in common. FEBS Lett. 1993 Apr 19;321(1):15–18. doi: 10.1016/0014-5793(93)80611-w. [DOI] [PubMed] [Google Scholar]
- Kato I., Noda M. ADP-ribosylation of cell membrane proteins by staphylococcal alpha-toxin and leukocidin in rabbit erythrocytes and polymorphonuclear leukocytes. FEBS Lett. 1989 Sep 11;255(1):59–62. doi: 10.1016/0014-5793(89)81060-9. [DOI] [PubMed] [Google Scholar]
- Kornblum J. S., Projan S. J., Moghazeh S. L., Novick R. P. A rapid method to quantitate non-labeled RNA species in bacterial cells. Gene. 1988;63(1):75–85. doi: 10.1016/0378-1119(88)90547-1. [DOI] [PubMed] [Google Scholar]
- König B., Köller M., Prevost G., Piemont Y., Alouf J. E., Schreiner A., König W. Activation of human effector cells by different bacterial toxins (leukocidin, alveolysin, and erythrogenic toxin A): generation of interleukin-8. Infect Immun. 1994 Nov;62(11):4831–4837. doi: 10.1128/iai.62.11.4831-4837.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Noda M., Hirayama T., Kato I., Matsuda F. Crystallization and properties of staphylococcal leukocidin. Biochim Biophys Acta. 1980 Nov 17;633(1):33–44. doi: 10.1016/0304-4165(80)90035-5. [DOI] [PubMed] [Google Scholar]
- Noda M., Kato I., Hirayama T., Matsuda F. Fixation and inactivation of staphylococcal leukocidin by phosphatidylcholine and ganglioside GM1 in rabbit polymorphonuclear leukocytes. Infect Immun. 1980 Aug;29(2):678–684. doi: 10.1128/iai.29.2.678-684.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Noda M., Kato I., Matsuda F., Hirayama T. Mode of action of staphylococcal leukocidin: relationship between binding of 125I-labeled S and F components of leukocidin to rabbit polymorphonuclear leukocytes and leukocidin activity. Infect Immun. 1981 Nov;34(2):362–367. doi: 10.1128/iai.34.2.362-367.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Plommet M. Preparation and purification of gamma-hemolysin of staphylococci. Methods Enzymol. 1988;165:8–16. doi: 10.1016/s0076-6879(88)65005-1. [DOI] [PubMed] [Google Scholar]
- Prevost G., Couppie P., Prevost P., Gayet S., Petiau P., Cribier B., Monteil H., Piemont Y. Epidemiological data on Staphylococcus aureus strains producing synergohymenotropic toxins. J Med Microbiol. 1995 Apr;42(4):237–245. doi: 10.1099/00222615-42-4-237. [DOI] [PubMed] [Google Scholar]
- Rahman A., Izaki K., Kamio Y. Gamma-hemolysin genes in the same family with lukF and lukS genes in methicillin resistant Staphylococcus aureus. Biosci Biotechnol Biochem. 1993 Jul;57(7):1234–1236. doi: 10.1271/bbb.57.1234. [DOI] [PubMed] [Google Scholar]
- Rahman A., Nariya H., Izaki K., Kato I., Kamio Y. Molecular cloning and nucleotide sequence of leukocidin F-component gene (lukF) from methicillin resistant Staphylococcus aureus. Biochem Biophys Res Commun. 1992 Apr 30;184(2):640–646. doi: 10.1016/0006-291x(92)90637-z. [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]
- Soboll H., Ito A., Schaeg W., Blobel H. Leukozidin von Staphylokokken verschiedener Herkunft. Zentralbl Bakteriol Orig A. 1973 Jul;224(2):184–193. [PubMed] [Google Scholar]
- Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
- Supersac G., Prevost G., Piemont Y. Sequencing of leucocidin R from Staphylococcus aureus P83 suggests that staphylococcal leucocidins and gamma-hemolysin are members of a single, two-component family of toxins. Infect Immun. 1993 Feb;61(2):580–587. doi: 10.1128/iai.61.2.580-587.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor A. G., Bernheimer A. W. Further characterization of staphylococcal gamma-hemolysin. Infect Immun. 1974 Jul;10(1):54–59. doi: 10.1128/iai.10.1.54-59.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [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]
- WOODIN A. M. Purification of the two components of leucocidin from Staphylococcus aureus. Biochem J. 1960 Apr;75:158–165. doi: 10.1042/bj0750158. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ward P. D., Turner W. H. Identification of staphylococcal Panton-Valentine leukocidin as a potent dermonecrotic toxin. Infect Immun. 1980 May;28(2):393–397. doi: 10.1128/iai.28.2.393-397.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woodin A. M., Wieneke A. A. Role of leucocidin and triphosphoinositide in the control of potassium permeability. Nature. 1968 Oct 19;220(5164):283–286. doi: 10.1038/220283a0. [DOI] [PubMed] [Google Scholar]
- Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
- von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]