TABLE 1.
Bacterial strains and plasmids used in this study
| Strain/plasmid | Characteristics | Source/Ref. |
|---|---|---|
| EPEC E2348/69 | Wild type EPEC O127:H6 | 23 |
| ΔnleE | EPEC E2348/69 ΔnleE cmR | 18 |
| ΔPP4/IE6 | EPEC E2348/69 ΔPP4/IE6 double island deletion | 6 |
| S. flexneri 6 | Wild type S. flexneri 6 | 6 |
| ΔospZ | S. flexneri 6 ΔospZ kanR | This study |
| S. cerevisiae | S. cerevisiae AH109 | Clontech |
| pTrc99a | Cloning vector for expression of proteins from Ptrc | GE Healthcare |
| pNleE | nleE from EPEC E2348/69 in pTrc99a | 6 |
| pNleEΔ49–52 | nleE from EPEC E2348/69 carrying the GITR deletion in pTrc99a | This study |
| pNleE49AAAA52 | nleE from EPEC E2348/69 carrying alanine substitution for each amino acid in the GITR motif in pTrc99a | This study |
| pEGFP-C2 | GFP expression vector | Clontech |
| pEGFP-NleE | nleE from EPEC E2348/69 in pTrc99a | 18 |
| pEGFP-NleE1–53 | Truncation of nleE from EPEC E2348/69 encoding amino acids 1–53 in pEGFP-C2 | This study |
| pEGFP-NleEΔ34–52 | Full-length nleE from EPEC E2348/69 carrying the GITR deletion in pEGFP-C2 | This study |
| pEGFP-NleE49AAAA52 | nleE from EPEC E2348/69 carrying alanine substitution for each amino acid in the GITR motif in pEGFP-C2 | This study |
| pEGFP-OspZ | ospZ from S. flexneri 6 in pEGFP-C2 | This study |
| pEGFP-OspZ49AAAA52 | ospZ from S. flexneri 6 carrying alanine substitution for each amino acid in the GITR motif in pEGFP-C2 | This study |
| pGEX-6P-1 | GST expression vector | GE Healthcare |
| pEGFP-NleE34–52 | The region between amino acids 34 and 52 of nleE in pEGFP-C2 | This study |
| pGST-NleE34–52 | The region between amino acids 34 and 52 of nleE in pGEX-6P-1 | This study |
| pGST-NleE | nleE from EPEC E2348/69 in pGEX-6P-1 | This study |
| pGST-NleE49AAAA52 | Full-length nleE from EPEC E2348/69 carrying a mutation in the motif 49GITR52 in pGEX-6P-1 | This study |
| pGST-OspZ | ospZ from S. flexneri 6 in pGEX-6P-1 | This study |
| pGST-OspZ49AAAA52 | Full-length ospZ from S. flexneri 6 carrying a mutation in the motif 49GITR52 in pGEX-6P-1 | This study |
| pGBKT7 | Yeast expression vector | Clontech |
| pGBKT7-NleE | nleE from EPEC E2348/69 in pGBKT7 | This study |
| pGBKT7-NleE1–53 | Truncation of nleE from EPEC E2348/69 encoding amino acids 1–53 in pGBKT7 | This study |
| pGBKT7-NleE1–82 | Truncation of nleE from EPEC E2348/69 encoding amino acids 1–82 in pGBKT7 | This study |
| pGBKT7-NleE1–118 | Truncation of nleE from EPEC E2348/69 encoding amino acids 1–118 in pGBKT7 | This study |
| pGBKT7-NleE54–224 | Truncation of nleE from EPEC E2348/69 encoding amino acids 54–224 in pGBKT7 | This study |
| pGBKT7-NleE83–224 | Truncation of nleE from EPEC E2348/69 encoding amino acids 83–224 in pGBKT7 | This study |
| pGBKT7-NleE119–224 | Truncation of nleE from EPEC E2348/69 encoding amino acids 119–224 in pGBKT7 | This study |
| pGBKT7-NleE6A | nleE from EPEC E2348/69 in pGBKT7 carrying alanine substitutions for each amino acid in the IDSYMK motif | This study |
| pGBKT7-NleE34–52 | The region between amino acids 34 and 52 of nleE in pGBKT7 | This study |
| pGBKT7-NleEΔ34–52 | Full-length nleE from EPEC E2348/69 carrying the deletion between amino acids 34 and 52 in pGBKT7 | This study |
| pGBKT7-NleEΔ41–52 | Full-length nleE from EPEC E2348/69 carrying the deletion between amino acids 41 and 52 in pGBKT7 | This study |
| pGBKT7-NleEΔ45–52 | Full-length nleE from EPEC E2348/69 carrying the deletion between amino acids 45 and 52 in pGBKT7 | This study |
| pGBKT7-NleEΔ49–52 | Full-length nleE from EPEC E2348/69 carrying the GITR deletion in pGBKT7 | This study |
| pGBKT7-OspZ | Full-length ospZ from S. flexneri 6 in pGBKT7 | This study |
| pCX340 | Cloning vector for constructing TEM-1 β-lactamase fusions | 20 |
| pCX340-NleE | nleE from EPEC E2348/69 in pCX340 | This study |
| pCX340-NleE54–224 | Truncation of nleE from EPEC E2348/69 encoding amino acids 54–224 in pCX340 | This study |
| pCX340-NleEΔ34–52 | Full-length nleE from EPEC E2348/69 carrying a deletion of amino acids between 34 and 52 in pCX340 | This study |
| pCX340-NleEΔ49–52 | Full-length nleE from EPEC E2348/69 carrying the GITR deletion in pCX340 | This study |
| pcDNA3 | Mammalian cell expression vector | Invitrogen |
| pcDNA3-TAB3–3×FLAG | Full-length human TAB3 with a C-terminal 3 × FLAG tag in pcDNA3 | This study |
| pcDNA3-TAB2–3×FLAG | Full-length human TAB3 with a C-terminal 3 × FLAG tag in pcDNA3 | This study |
| pcDNA3-ZRANB3–3×FLAG | Full-length human ZRANB3 with a C-terminal 3 × FLAG tag in pcDNA3 | This study |
| pGADT7 | Yeast expression vector | Clontech |
| pGADT7-TAB3 | Full-length human TAB3 in pGADT7 | This study |
| pRL-TK | Renilla luciferase vector | Promega |
| pNF-κB-Luc | Vector for measuring NF-κB-dependent luciferase expression | Clontech |