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https://hdl.handle.net/2440/51854
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Type: | Journal article |
Title: | Coiled-coil regions play 4 role in the function of the Shigella flexneri O-antigen chain length regulator Wzz(pHS2) |
Author: | Purins, L. Van Den Bosch, L. Richardson, V. Morona, R. |
Citation: | Microbiology, 2008; 154(Part 4):1104-1116 |
Publisher: | Soc General Microbiology |
Issue Date: | 2008 |
ISSN: | 1350-0872 1465-2080 |
Statement of Responsibility: | Leanne Purins, Luisa Van Den Bosch, Vanessa Richardson and Renato Morona |
Abstract: | Regulation of the length of the O-antigen (Oag) chain attached to LPS in Shigella flexneri is important for virulence and is dependent on the inner-membrane protein Wzz. A lack of high-resolution structural data for Wzz has hampered efforts so far to correlate mutations affecting function of Wzz with structure and describe a mechanism for chain length regulation. Here we have used secondary structure prediction to show that the periplasmic domain of the Wzz(pHS2) protein has three regions of significant coiled-coil (CC) potential, two of which lie within an extended helical region. We describe here the first site-directed mutagenesis study to investigate the role of individual predicted CC regions (CCRs) in Wzz function and oligomerization. We found that CCRs 2 and 3 are necessary for wild-type Oag chain length regulation by Wzz(pHS2). The in vivo cross-linking profile of mutants affected in the three CCRs was not altered, indicating that individually each CCR is not required for oligomerization. Interestingly, the CCR3 mutation resulted in a temperature-sensitive phenotype and an inhibitory effect on Oag polymerization. Analysis of Wzz(pHS2) and the mutant constructs in a S. flexneri degP mutant showed that DegP did not affect the function of wild-type Wzz(pHS2) but its presence influenced the phenotype of the Wzz(pHS2) CCR3 mutant. Additionally, the phenotype of the Wzz(pHS2) CCR3 mutant was suppressed by a cis mutation near the putative cytoplasmic C-terminus of Wzz(pHS2). |
Keywords: | Shigella flexneri Serine Endopeptidases O Antigens Periplasmic Proteins Bacterial Proteins Heat-Shock Proteins Mutagenesis, Site-Directed Gene Deletion Suppression, Genetic Amino Acid Sequence Base Sequence Protein Structure, Secondary Molecular Sequence Data Hot Temperature |
DOI: | 10.1099/mic.0.2007/014225-0 |
Published version: | http://dx.doi.org/10.1099/mic.0.2007/014225-0 |
Appears in Collections: | Aurora harvest Molecular and Biomedical Science publications |
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