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Deletion of Rho1Gene and Preliminary Characterization on Its Function in Aspergillus Fumigatus

Author: HanGaiGe
Tutor: HanLi
School: Central South University
Course: Biology
Keywords: Aspergillus fumigatus Rho1 Gene deletion Cell wallintegrity Invasion
CLC: R379
Type: Master's thesis
Year: 2013
Downloads: 9
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Abstract


Abstract:As one of the most important fungal pathogens, Aspergillus fumigatus causes invasive aspergillosis with very high mortality. Rho1is a regulatory subunit of β-1,3-D-glucan synthase that generate cell wall β-1,3-D-glucans. However, there is quite limited data about the function of rhol gene in A. fumigatus due to the absence of rhol gene-null mutant of A. fumigatus. In this study the rhol-deleted strain(Δrhol) and the rhol-gene complemented strain (rho1C) of A. fumigatus AF293were successfully constructed with homologous recombination. It was found that Arhol was not able to grow at37℃, which was quite different from the rho1-gene-deleted strains in other Aspergillus. The Arhol mutant could grow at30℃but its radial growth rate was much lower than that of wild-type strain; in contrast, it grew a little bit faster than wild-type strain at25℃. The edge of the colony of Arhol mutant is smoother and the conidiation was also significantly less than that of wild-type strain. The hyphal tip became thinner and the tip complex was lost with the rhol gene deletion. The resistance of Arhol against high-concentration cell wall perturbing agents and oxidative stress was stronger than the wild-type strain; however, no significant change on the susceptibility to the osmotic pressure and alkaline environment (high pH value) was found in Arhol mutant. In addition, the deletion of rhol gene promoted the production of ROS inside A. fumigatus and this increase was obviously inhibited by the treatment by cell wall perturbing agents. Further, Arhol mutant adhered to the lung II A549epithelial cells and induced the production of ROS in the host cells as efficient as wild-type strain; however, invasion of Arhol conidia into A549cells was dramatically reduced compared to the wild-type strain. In the all above observations, the characterization of rho1C showed very high similarity with that of wild-type strain. Taken together, the rhol gene may be involved in the regulation of temperature tolerance, cell wall integrity and oxidative stress response and internalization into host cells of A. fumigatus. More investigation is necessary to elucidate its detailed regulation mechanism.

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CLC: > Medicine, health > Basic Medical > Medical Microbiology ( pathogenic bacteriology,pathogenic microbiology ) > Pathogenic fungi ( molds) and actinomycetes
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