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Biochem Lit - Essay Example

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In their paper, “Differential Roles of the ChiB Chitinase in Autolysis and Cell Death of Aspergillus nidulans” Kwang-Soo Shin and co-workers explored the mechanism controlling autolysis , a natural process of enzymatic self degradation of cells. FlbA is a regulator of G…
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In their paper, “Differential Roles of the ChiB Chitinase in Autolysis and Cell Death of Aspergillus nidulans” Kwang-Soo Shin and co-workers explored the mechanism controlling autolysis , a natural process of enzymatic self degradation of cells. FlbA is a regulator of G protein signaling. In the absence of this regulator, autolysis becomes a predominant phenomenon. This was confirmed by flbA deletion (∆flbA) mutant which exhibited uncontrolled proliferation of hyphal mass, escaped sporulation and eventually resulted in autolysis in previous studies. It was ascertained in this course of study that ChiB, a gene coding for endo Chitinase B, is in part responsible for induction of autolysis by flbA gene. Also though ChiB was found to be necessary for ∆flbA induced autolysis its overexpression did not accelerate or enhance hyphal disintegration thus suggesting that ChiB is necessary but not sufficient for autolysis in submerge-cultured A.nidulans. Alamar Blue reduction test which corresponds to the activity of the living cell mitochondria was employed to assess the dispensability of ChiB gene in autolysis. By AB reduction it was seen that both Wild Type strain and ChiB deletion ∆ChiB mutant showed the same death pattern from day 1 to day 8. Thus though ChiB was required for autolysis it was not totally essential. To study the proteins involved in autolysis, the proteins upregulated in ∆flbA and ∆flbA∆ChiB were studied and five proteins were found to be upregulated. These were identified as chitinase (ChiB), dipeptidyl peptidase V (DppV), O-glycosyl compound hydrolase, -N-acetylhexosaminidase (NagA), and myo-inositol-1-phosphate synthase (InoB) on SDS PAGE after Liquid chromatography-tandem mass spectroscopy. Off these 5 proteing NagA was speculated to be associated with cell death. This was because ∆NagA mutant showed a significantly higher percentage reduction in Alamar Blue as compared to other proteins on day 2 of incubation indicating more viable cells. The absence of InoB or AN2395.2 (a probable hydrolase) was seen to cause accelerated cell death and disorganization of mycelial pellets suggesting that these proteins might be needed for the controlled progression of cell death. FlbA gene negatively controls G protein mediated proliferation signaling likely by acting as GTPase activating protein for FadAGα. It was also concluded in other studies that FadA and SfaD signaling negatively regulate the cell wall chitin content and cell wall structure in A.nidulans. FadA was also found to mediate signaling involved in the initiation of apoptosis by another study. Finally, yet another study concluded that the FadA heterotrimeric G protein complex was found to be a part of the signal transduction pathway that promotes apoptosis in hyphae exposed to chemical farnesol. Thus on the same lines, the research in this paper lead to the formulation of a working model which suggested that FadA and SfaD were constitutively activated in the absence of regulating flbA gene which resulted in accelerated autolysis and that ChiB and NagA “ play differential roles in these interconnected, yet separate biological processes” as aptly summarized in Toto from the paper.
Work Cited
Shin, Kwang-Soo. Differential Roles of the ChiB Chitinase in Autolysis and Cell Death
of Aspergillus nidulans Republic of Korea. 2009. American Society of Microbiology Print. Read More
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