Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/88961
Citations | ||
Scopus | Web of Science® | Altmetric |
---|---|---|
?
|
?
|
Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.author | Whittall, J. | - |
dc.contributor.author | Morona, R. | - |
dc.contributor.author | Standish, A. | - |
dc.contributor.editor | de Boer, P. | - |
dc.date.issued | 2015 | - |
dc.identifier.citation | Journal of Bacteriology, 2015; 197(1):120-127 | - |
dc.identifier.issn | 1098-5530 | - |
dc.identifier.issn | 1098-5530 | - |
dc.identifier.uri | http://hdl.handle.net/2440/88961 | - |
dc.description.abstract | In Gram-positive bacteria, tyrosine kinases are split into two proteins, the cytoplasmic tyrosine kinase and a transmembrane adaptor protein. In Streptococcus pneumoniae this transmembrane adaptor is CpsC, with the C-terminus of CpsC critical for interaction and subsequent tyrosine kinase activity of CpsD. Topology predictions suggest CpsC has two transmembrane domains, with the N and C-termini present in the cytoplasm. In order to investigate CpsC topology, we used a chromosomal HA-tagged Cps2C protein in D39. Incubation of both protoplasts and membranes with the CP-B resulted in complete degradation of HA-Cps2C in all cases, indicating that the C-terminus of Cps2C was likely extra-cytoplasmic, and hence the protein's topology was not as predicted. Similar results were seen with membranes from TIGR4, indicating Cps4C also showed similar topology. A chromosomally encoded fusion of HA-Cps2C and Cps2D was not degraded by CP-B, suggesting that the fusion fixed the C-terminus within the cytoplasm. However, capsule synthesis was unaltered by this fusion. Detection of the CpsC C-terminus by flow cytometry indicated that it was extra-cytoplasmic in approximately 30% of cells. Interestingly, a mutant in the protein tyrosine phosphatase CpsB had a significantly greater proportion of positive cells, although this affect was independent of its phosphatase activity. Our data indicate that CpsC possesses a varied topology, with the C-terminus flipping across the cytoplasmic membrane where it interacts with CpsD in order to regulate tyrosine kinase activity. | - |
dc.description.statementofresponsibility | Jonathan J. Whittall, Renato Morona, Alistair J. Standish | - |
dc.language.iso | en | - |
dc.publisher | American Society for Microbiology | - |
dc.rights | Copyright © 2015, American Society for Microbiology. All Rights Reserved. | - |
dc.source.uri | http://dx.doi.org/10.1128/jb.02106-14 | - |
dc.subject | Streptococcus pneumoniae | - |
dc.subject | Polysaccharides, Bacterial | - |
dc.subject | Bacterial Proteins | - |
dc.subject | Virulence Factors | - |
dc.subject | Flow Cytometry | - |
dc.subject | Gene Expression Regulation, Bacterial | - |
dc.subject | Gene Expression Regulation, Enzymologic | - |
dc.subject | Protein Conformation | - |
dc.title | Topology of Streptococcus pneumoniae CpsC, a Polysaccharide Copolymerase and Bacterial Protein Tyrosine Kinase Adaptor Protein | - |
dc.type | Journal article | - |
dc.identifier.doi | 10.1128/jb.02106-14 | - |
dc.relation.grant | http://purl.org/au-research/grants/nhmrc/1048749 | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | Morona, R. [0000-0001-7009-7440] | - |
Appears in Collections: | Aurora harvest 7 Microbiology and Immunology publications |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
hdl_88961.pdf | Accepted version | 1.2 MB | Adobe PDF | View/Open |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.