Author Description

This dissertation, "Structural Studies of Two Nucleoid-associated Proteins: Histone-like Nucleoid-structuring Protein H-NS and Α-hemolysin Expression-modulating Protein Hha" by Wei, Cao,, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: In prokaryotic cells, the nucleoid contains almost all the genetic materials as well as a number of nucleoid structuring factors. The nucleoid-associated proteins (NAPs) are known to have low molecular weight and the ability to form dimer or oligomer, and most of them can bind to DNA for regulation of gene expression. The Histone-like nucleoid structuring protein H-NS, well studied as one of the NAPs, acts as a global transcriptional repressor. It has independent functional N-terminal domain for oligomerization and C-terminal domain for DNA binding, joined by a flexible linker. H-NS contributes to horizontal genes transfer and responses to environmental factors like temperature or pH, which would influence the oligomerization ability of H-NS and DNA binding. The α-hemolysin expression-modulating protein Hha is a member of the Hha-YmoA family, expressed only in Gram-negative Enterobacteriaceae as a modulator of virulence factors expression. In E. coli, the binding of Hha to H-NS can modulate the expression of α-hemolysin operon, which is essential for the H-NS-regulated gene expression. In this study, both Hha and the oligomerization domain of H-NS (H-NS64) were expressed in E. coli and the purified proteins were crystallized. The Hha crystals diffracted to 2.2 A; and the HhA/H-NS complex crystals diffracted to 1.8 A. Both structures were successfully determined by molecular