Identification of Pexophagy Genes by Restriction Enzyme-Mediated Integration

Schröder, Laura; Glick, Benjamin S.; Dunn, Billy

doi:10.1385/1-59745-456-7:203

Cited by 5 publications

(5 citation statements)

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“…However, methylotrophic yeasts have provided most of the important details regarding the selectivity of pexophagy, which is the primary focus of this mini-review. Specific genetic screens using UV, chemical or insertional mutagenesis strategies in yeasts have elucidated genes required for pexophagy [11][12][13][14][15][16][17]. Because selective autophagy pathways rely on components of the core autophagic machinery, these screens have revealed components of the core autophagy machinery as well as unique selectivity factors that adapt the core autophagy machinery for pexophagy [14,16].…”

Section: Model Systems Used To Examine Pexophagymentioning

confidence: 99%

Molecular mechanism and physiological role of pexophagy

et al. 2010

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a b s t r a c tPexophagy is a selective autophagy process wherein damaged and/or superfluous peroxisomes undergo vacuolar degradation. In methylotropic yeasts, where pexophagy has been studied most extensively, this process occurs by either micro-or macropexophagy: processes analogous to microand macroautophagy. Recent studies have identified specific factors and illustrated mechanisms involved in pexophagy. Although mechanistically pexophagy relies heavily on the core autophagic machinery, the latest findings about the role of auxiliary pexophagy factors have highlighted specialized membrane structures required for micropexophagy, and shown how cargo selectivity is achieved and how cargo size dictates the requirement for these factors during pexophagy. These insights and additional observations in the literature provide a framework for an understanding of the physiological role(s) of pexophagy.

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Section: Model Systems Used To Examine Pexophagymentioning

confidence: 99%

Molecular mechanism and physiological role of pexophagy

et al. 2010

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“…The enzyme extract was incubated with 10 U/mL horseradish peroxidase, 10 mmol/L PBS (pH 7.5), 1 mmol/L 4-aminoantipyrine, 4.3 mol/L phenol, and 200 mmol/L methanol at 37°C for 10 min. A red-colored product was obtained, which was evaluated using a spectrophotometer at 500 nm (722S, Shanghai Precision Scientific Instrument Co., Ltd, Shanghai, China) with the modification of Schroder’s method ( Schroder et al, 2007 ). One unit of AOX activity was defined as the amount of AOX required to produce 1 μmol H 2 O 2 per minute.…”

Section: Methodsmentioning

confidence: 99%

Energy Charge as an Indicator of Pexophagy in Pichia pastoris

Zhang

Liu

2017

Front. Microbiol.

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Pichia pastoris is a good model for pexophagy research owing to its diverse pexophagy modes (macropexophagy and micropexophagy) exhibited during carbon-source shift from methanol to other carbon sources. The critical condition that triggers activation of macropexophagy and micropexophagy is important for clarifying the P. pastoris pexophagy mechanism and human peroxisomal disorders. In this study, the pexophagy modes of P. pastoris were confirmed by green fluorescent protein expression and alcohol oxidase and formate dehydrogenase activities. Furthermore, intracellular energy charge (EC) was found to be a determinant of pexophagy activation. During methanol induction, the EC was about 0.5. And the final EC value was related to the pexophagy mode when carbon source switched from methanol to others. Macropexophagy and micropexophagy occurred when the EC increased to 0.6–0.75 and above 0.75, respectively. Thus, different EC values were considered as the important factor to trigger different pexophagy modes in P. pastoris. The results obtained in this study could help in achieving better control of the pexophagy modes to study the pexophagy mechanism.

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“…Briefly, cells were harvested in late-log phase and collected by centrifugation. The pellet was washed with sterile distilled water and resuspended in an appropriate volume of lysis buffer (100 mM Tris-HCl (pH 8.0), 20 g l -1 Triton X-100, 10 g l -1 sodium dodecyl sulfate, 100 mM NaCl, 10 mM Na2EDTA) (Schroder et al, 2007). For cell lysis, resuspended cells were vortexed vigorously for one minute and then kept at 65 °C in a water bath for 20 minutes, after which they were again vortexed for one minute.…”

Section: Genomic Dna Preparationmentioning

confidence: 99%