Reference no: EM132927569
Questions and answers on paper by Besteiro et al., (2006) Endosome sorting and autophagy are essential for differentiation and virulence of Leishmania major. J. Biol. Chem. 281, 11384-11396.
1) What does ESCRT stand for?
2) Draw a diagram that shows the main stages of the basic mechanism for sorting lysosomal proteins in eukaryotes. What is its relevance for Leishmania virulence?
3) Briefly define autophagy and mention two main roles that have been suggested for this process in eukaryotic organisms.
4) Which is the Leishmania life cycle differentiation step studied by the authors and in which natural host does it take place?
5) What does LmjVPS4E235Q stand for and what is its purpose in the study?
6) Which of the following statements is the most likely explanation for the use of wortmannin and methyladenine in the starvation induced autophagy assay in the study?
Justify your choice of statement.
7) Briefly explain the purpose of using heat-inactivated serum in the complement-mediated lysis tests used in the study.
8) Briefly explain how the MTT assay measures cell viability.
9) How were the data from macrophage infections and MTT assays analysed for significance by the authors?
10) Briefly explain the in vitro growth profiles shown by Leishmania over-expressing VPS4E235Q in Figure 1.
11) Is LmjVPS4E235Q defective in metacyclogenesis? Justify your answer using data shown in Panels B, C and D of Figure 4.
12) Is Atg8 a valid marker for monitoring autophagosome formation in Leishmania? Justify your answer including data shown in Panels B and C of Figure 6.
13) What can you conclude about Leishmania survival and autophagosome formation under starvation conditions in the VPS4E235Q mutant Leishmania? Justify your answer using data shown in Panels D and E of Figure 7.
14) How, according to the authors, might susceptibility to starvation conditions and metacyclogenesis be related in both the VPS4E235Q and the ATG4.2 mutant Leishmania?