A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved Sequence
ABSTRACT Many Gram-negative bacteria utilize specialized secretion systems to inject proteins (effectors) directly into host cells. Little is known regarding how bacteria ensure that only small subsets of the thousands of proteins they encode are recognized as substrates of the secretion systems, li...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2012-03-01
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| Series: | mBio |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.00243-11 |
| _version_ | 1818424729442189312 |
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| author | Sonia C. P. Costa Alexa M. Schmitz Fathima F. Jahufar Justin D. Boyd Min Y. Cho Marcie A. Glicksman Cammie F. Lesser |
| author_facet | Sonia C. P. Costa Alexa M. Schmitz Fathima F. Jahufar Justin D. Boyd Min Y. Cho Marcie A. Glicksman Cammie F. Lesser |
| author_sort | Sonia C. P. Costa |
| collection | DOAJ |
| description | ABSTRACT Many Gram-negative bacteria utilize specialized secretion systems to inject proteins (effectors) directly into host cells. Little is known regarding how bacteria ensure that only small subsets of the thousands of proteins they encode are recognized as substrates of the secretion systems, limiting their identification through bioinformatic analyses. Many of these proteins require chaperones to direct their secretion. Here, using the newly described protein interaction platform assay, we demonstrate that type 3 secretion system class IB chaperones from one bacterium directly bind their own effectors as well as those from other species. In addition, we observe that expression of class IB homologs from seven species, including pathogens and endosymbionts, mediate the translocation of effectors from Shigella directly into host cells, demonstrating that class IB chaperones are often functionally interchangeable. Notably, class IB chaperones bind numerous effectors. However, as previously proposed, they are not promiscuous; rather they recognize a defined sequence that we designate the conserved chaperone-binding domain (CCBD) sequence [(LMIF)1XXX(IV)5XX(IV)8X(N)10]. This sequence is the first defined amino acid sequence to be identified for any interspecies bacterial secretion system, i.e., a system that delivers proteins directly into eukaryotic cells. This sequence provides a new means to identify substrates of type III secretion systems. Indeed, using a pattern search algorithm for the CCBD sequence, we have identified the first two probable effectors from an endosymbiont, Sodalis glossinidius. IMPORTANCE Many Gram-negative pathogens utilize type 3 secretion systems to deliver tens of effectors into host cells. In order to understand the diverse ways that these organisms cause disease, it is necessary to identify their effectors, many of which require chaperones to be secreted. Here we establish that class IB chaperones are not promiscuous, as previously proposed, but rather recognize a conserved effector sequence. We demonstrate that pattern search algorithms based on this defined sequence can be used to identify previously unknown effectors. Furthermore, we observe that class IB chaperones from at least seven bacterial species are functionally interchangeable. Not only do they bind and mediate the delivery of their own set of effectors into host cells but they also bind to type 3 substrates from other bacteria, suggesting that inhibitors that block chaperone-effector interactions could provide a novel means to effectively treat infections due to Gram-negative pathogens, including organisms resistant to currently available antibiotics. |
| first_indexed | 2024-12-14T14:02:40Z |
| format | Article |
| id | doaj.art-a5b66c4311bb43ce8b399674eac47930 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-14T14:02:40Z |
| publishDate | 2012-03-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-a5b66c4311bb43ce8b399674eac479302022-12-21T22:58:42ZengAmerican Society for MicrobiologymBio2150-75112012-03-013110.1128/mBio.00243-11A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved SequenceSonia C. P. Costa0Alexa M. Schmitz1Fathima F. Jahufar2Justin D. Boyd3Min Y. Cho4Marcie A. Glicksman5Cammie F. Lesser6Division of Infectious Diseases, Department of Medicine (Microbiology and Molecular Genetics), Massachusetts General Hospital and Harvard Medical School, Cambridge, Massachusetts, USADivision of Infectious Diseases, Department of Medicine (Microbiology and Molecular Genetics), Massachusetts General Hospital and Harvard Medical School, Cambridge, Massachusetts, USADivision of Infectious Diseases, Department of Medicine (Microbiology and Molecular Genetics), Massachusetts General Hospital and Harvard Medical School, Cambridge, Massachusetts, USALaboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women’s Hospital, and Harvard Medical School, Cambridge, Massachusetts, USADivision of Infectious Diseases, Department of Medicine (Microbiology and Molecular Genetics), Massachusetts General Hospital and Harvard Medical School, Cambridge, Massachusetts, USALaboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women’s Hospital, and Harvard Medical School, Cambridge, Massachusetts, USADivision of Infectious Diseases, Department of Medicine (Microbiology and Molecular Genetics), Massachusetts General Hospital and Harvard Medical School, Cambridge, Massachusetts, USAABSTRACT Many Gram-negative bacteria utilize specialized secretion systems to inject proteins (effectors) directly into host cells. Little is known regarding how bacteria ensure that only small subsets of the thousands of proteins they encode are recognized as substrates of the secretion systems, limiting their identification through bioinformatic analyses. Many of these proteins require chaperones to direct their secretion. Here, using the newly described protein interaction platform assay, we demonstrate that type 3 secretion system class IB chaperones from one bacterium directly bind their own effectors as well as those from other species. In addition, we observe that expression of class IB homologs from seven species, including pathogens and endosymbionts, mediate the translocation of effectors from Shigella directly into host cells, demonstrating that class IB chaperones are often functionally interchangeable. Notably, class IB chaperones bind numerous effectors. However, as previously proposed, they are not promiscuous; rather they recognize a defined sequence that we designate the conserved chaperone-binding domain (CCBD) sequence [(LMIF)1XXX(IV)5XX(IV)8X(N)10]. This sequence is the first defined amino acid sequence to be identified for any interspecies bacterial secretion system, i.e., a system that delivers proteins directly into eukaryotic cells. This sequence provides a new means to identify substrates of type III secretion systems. Indeed, using a pattern search algorithm for the CCBD sequence, we have identified the first two probable effectors from an endosymbiont, Sodalis glossinidius. IMPORTANCE Many Gram-negative pathogens utilize type 3 secretion systems to deliver tens of effectors into host cells. In order to understand the diverse ways that these organisms cause disease, it is necessary to identify their effectors, many of which require chaperones to be secreted. Here we establish that class IB chaperones are not promiscuous, as previously proposed, but rather recognize a conserved effector sequence. We demonstrate that pattern search algorithms based on this defined sequence can be used to identify previously unknown effectors. Furthermore, we observe that class IB chaperones from at least seven bacterial species are functionally interchangeable. Not only do they bind and mediate the delivery of their own set of effectors into host cells but they also bind to type 3 substrates from other bacteria, suggesting that inhibitors that block chaperone-effector interactions could provide a novel means to effectively treat infections due to Gram-negative pathogens, including organisms resistant to currently available antibiotics.https://journals.asm.org/doi/10.1128/mBio.00243-11 |
| spellingShingle | Sonia C. P. Costa Alexa M. Schmitz Fathima F. Jahufar Justin D. Boyd Min Y. Cho Marcie A. Glicksman Cammie F. Lesser A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved Sequence mBio |
| title | A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved Sequence |
| title_full | A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved Sequence |
| title_fullStr | A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved Sequence |
| title_full_unstemmed | A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved Sequence |
| title_short | A New Means To Identify Type 3 Secreted Effectors: Functionally Interchangeable Class IB Chaperones Recognize a Conserved Sequence |
| title_sort | new means to identify type 3 secreted effectors functionally interchangeable class ib chaperones recognize a conserved sequence |
| url | https://journals.asm.org/doi/10.1128/mBio.00243-11 |
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