Probing for a ribosome code in Escherichia coli using integration host factor
Integration host factor (IHF) is an important DNA binding and bending protein which regulates expression of a large number of genes in Escherichia coli. Our experimental results show that a polyhistidine-tagged single-chain IHF protein, sc1HF2, has the ability to interact with E. coli ribosomes in a...
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| Format: | Theses and Dissertations |
| Language: | English |
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Nanyang Technological University
2014
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| Online Access: | http://hdl.handle.net/10356/59963 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | Integration host factor (IHF) is an important DNA binding and bending protein which regulates expression of a large number of genes in Escherichia coli. Our experimental results show that a polyhistidine-tagged single-chain IHF protein, sc1HF2, has the ability to interact with E. coli ribosomes in a stable manner. To date, the interaction between IHF and ribosome has not been studied. Using sclHF2 we were able to purify ribosomes incorporated with sclHF2 and further confirmed the interaction takes place on the 50S ribosomal subunit. Moreover, we found correlations between IHF expression and bacterial survival mechanisms in times of starvation, especially ribosome
integrity. With the help of mass-spectrometry-based protein identification, we identified several binding partners exclusive for IHF-bound ribosomes. These findings suggest new roles for IHF in protein transport, stationary phase, ribosome metabolism and gene regulation. |
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