Open Access Highly Accessed Research article

Hyperexpression of α-hemolysin explains enhanced virulence of sequence type 93 community-associated methicillin-resistant Staphylococcus aureus

Kyra YL Chua1234, Ian R Monk1, Ya-Hsun Lin1, Torsten Seemann5, Kellie L Tuck6, Jessica L Porter1, Justin Stepnell1, Geoffrey W Coombs78, John K Davies2, Timothy P Stinear12 and Benjamin P Howden1234*

Author Affiliations

1 Department of Microbiology and Immunology, University of Melbourne, Melbourne, Victoria 3052, Australia

2 Department of Microbiology, Monash University, Clayton, Victoria 3800, Australia

3 Austin Centre for Infection Research (ACIR), Infectious Diseases Department, Austin Health, PO Box 5555, Heidelberg, Victoria 3084, Australia

4 Microbiology Department, Austin Health, Heidelberg, Victoria 3084, Australia

5 Victorian Bioinformatics Consortium, Monash University, Clayton, Victoria 3800, Australia

6 School of Chemistry, Monash University, Clayton, Victoria 3800, Australia

7 Australian Collaborating Centre for Enterococcus and Staphylococcus Species (ACCESS) Typing and Research, PathWest Laboratory Medicine-WA, Royal Perth Hospital, Perth, Western Australia 6000, Australia

8 School of Biomedical Sciences, Curtin University, Bentley, Western Australia 6102, Australia

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BMC Microbiology 2014, 14:31  doi:10.1186/1471-2180-14-31

Published: 10 February 2014

Additional files

Additional file 1:

Staphylococcus aureus ST93 strains used in this study.

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Additional file 2:

Expression of PSMα3 by ST93 strains and USA300. (A) Expression of deformylated PSMα3. (B) Expression of N-formylated PSMα3. Data shown are mean concentration (μg/ml) and SEM.

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Additional file 3:

Expression of Hla by ST93 strains and USA300. Hla expression measured by quantitative Western blot. Data shown are mean intensity of bands in arbitrary units and SEM.

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Additional file 4:

Hla Western Blot of JKD6159, JKD6159∆hla and JKD6159∆hla r (A) Western Blot demonstrating that JKD6159∆hla does not express Hla by Western Blot and that complementation of this mutant (JKD6159∆hla r) results in restoration of Hla expression. (B) Arrangement of PCR primers used PCR screen of JKD6159∆hla and JKD6159∆hla r. (C) PCR screen of 25 randomly selected S. aureus colonies obtained from two mice (mouse 4 and mouse 7) post skin infection with JKD6159∆hla r. The PCR primers used flank the region deleted in hla for the mutant and show incomplete penetration of the bacterial population with the repaired version of hla (17/25 with an intact allele for mouse 4 and 21/25 for mouse 7), thereby explaining the inability of the repaired mutant to fully restore the virulence phenotype in this infection model.

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Additional file 5:

Detection of formylated PSMα3 in JKD6159, JKD6159∆psmα and JKD6159∆psmα r by HPLC of culture filtrates. JKD6159∆psmα did not produce formylated PSMα3. Complementation of this strain resulted in restoration of formylated PSMα3 expression. In all strains δ-toxin expression was maintained.

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Additional file 6:

LukF-PV Western Blot of JKD6159 and JKD6159∆lukSF-PV. Western Blot demonstrating that JKD6159∆lukSF-PV does not express LukF-PV.

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Additional file 7:

Table of de novo assembly characteristics for S. aureus strains TPS3104, TPS3105 and TPS3106.

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Additional file 8:

Table of single nucleotide differences between JKD6159 and TPS3104.

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Additional file 9:

Table of single nucleotide differences between JKD6159 and TPS3105.

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Additional file 10:

Table of single nucleotide differences between JKD6159 and TPS3106.

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Additional file 11:

Table of primers used in this study.

Format: DOCX Size: 16KB Download file

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